File

File > New

Creates a plain single colored image. The dialog values (width, height, colors, depth, etc.) are automatically pre-filled according to properties of the current, or the most recent image.

Name

The filename text will be used as a image name.

Width, Height

Width and height of the new image in pixels.

Red, Green, Blue, Value

Intensity of all pixels in the new image. For an RGB image, define intensities of Red, Green and Blue channel. If you create a gray image, define its intensity (Value). The intensity range depends on the selected Depth: 0-255 (8bit), 0-1023 (10bit), 0-4095(12bit), or 0-65535(16bit).

Colors

Choose between an RGB image and a Gray image.

Depth

Bit depth (per channel) of the new image can be selected. Select 8, 10, 12 or 16.

File > Open

With this command, you can load one or more images to the application. A dialog box appears. Multiple files selection can be performed holding the Ctrl or the Shift key.

Look In

The last-used directory is displayed. Roll the combo box down to browse network neighborhood, network or local drives, etc.

Use current calibration

Some types of image files (bmp, jpg...) do not contain the calibration information. If the option Use current calibration is checked, such image isn't marked as uncalibrated after opening, but the current system calibration will be assigned to it.

Information

Image properties of the selected file can be displayed by pressing this button.

File name

Type in this edit box and you will be offered files which match the already-typed characters.

Files of type

Select an image format or All Images. Only files of the selected format will be displayed in the selection window.

Dims

Size of the selected image in pixels

Type

Number of channels and their bit depth.

ND

Number of dimensions (displayed for ND files only).

Size

Size of the file on disk.

Note

Time Step Definition: Some ND2 files do not have the time step defined properly. When you open such ND2 file, a window appears, where the proper time step between the frames shall be defined.

See Opening Image Files, Supported File Formats.

File > Save

Saves an image to a disk. In case of a new image that hasn't been saved yet, the File > Save As dialog box appears. Otherwise, the current version of the existing file is updated.

File > Save As

Saves the existing image under a different name. The following dialog box appears also when you invoke the Save command on a newly created image.

File name

Name of the image being saved.

Save as type

Specifies the file type, in which the image will be saved.

See also TIF Saving Options, AVI/MP4 Saving Options, Supported File Formats.

Save

When pushing Save, the Archive information dialog box appears (For LIM and JPEG2000 formats only). Confirm it to save the image.

Cancel

Closes the Save As Image dialog window without saving the image.

Compression

Choose the image compression ratio. Available compression modes depend on the selected Save as type.

More Options

Reveals more saving options.

Save Color Image

Check this option to save the color image into the file.

Save Binary Image

Check this option to save the binary layer into the file. If the selected file format does not support layers, the option turns in Burn Binary Image. The binary data are burned into the color data before saving the file.

Save Annotations

Check this option to save the annotation layer and the scale layer into the file. If the selected file format does not support layers, the option turns in Burn Annotations. The currently displayed annotation/scale data are burned into the color image before saving the file.

Read Only

Sets the Windows Read-only attribute to the file being saved.

TIF Saving Options

Keep Original Channel Combination

If checked, channel combination of the source image is preserved.

Bit Depth

Original bit depth of the new file can be maintained, or the file can be rescaled to 8- or 16-bits.

LUTs

LUTs of the source image can be ignored (None), automatic LUTs can be applied (Apply auto LUTs) or LUTs saved with the image can be applied (Apply saved LUTs).

Save Color Channel Data per Pixel

Color channel data can be saved per pixel to improve the TIFF file compatibility.

Mono Image for Each Channel

Each channel will be used to create a monochrome image.

RGB Image for Each Channel

Each channel will be used to create an RGB image in channel color or in grayscale.

Burn Scale

Inserts scale into the final image. The scale becomes part of the image and cannot be removed.

Burn Binary

Inserts the binary data into the final image. The binary data become part of the image and cannot be removed.

Burn Annotations

Inserts annotations into the final image. The annotations become part of the image and cannot be removed.

All Channels Merged to RGB Overlay Image

One RGB image is created from all channels available in the source image.

Note

Bit depth conversion cannot be used with spectral images.

Save OME Metadata

OME metadata can be saved with the TIFF file to increase the TIFF file compatibility (e.g. proper channel designation).

See also Supported File Formats.

AVI/MP4 Saving Options

AVI and MP4 movie files can be created from a ND2 file. When the *.AVI or *.MP4 format is selected in the Save as type combo box, the following options appear:

Compression

Select which video compression is applied on created movie files. See About Video Compression.

XY Points

When the ND document contains XY points, you can decide how this dimension is saved. Either only the current point can be saved into the file, or all XY points can be saved to separate files which are recognizable by the "_number" suffix.

Output Timing Parameters

Timing of frames of the resulting movie may equal the Original Acquisition Time. If this does not suit your needs, define a custom interval in ms.

Experiment Loop

Select the dimension, frames of which will be used to create the movie. The dimension must be present in the ND2 file necessarily.

Use Time Loop

The T dimension frames of the current Z stack position will be used.

Use Z-stack Loop

The Z dimension frames of the current Time position will be used.

Use Z-stack and Time Loop

All frames of the ND2 file will be used in the movie.

When the Multipoint (XY) dimension is present, only frames of the current position are used automatically.

File > Save/Export to TIFF Files

Opens the Export Images to TIFF dialog window enabling to export image files into TIFF format and to define its target parameters.

Dialog Window Options

Image Info

Displays basic information about the currently selected image file together with its preview.

Current Document, All Opened Documents, Multiple Files

Determines which images will be exported to TIFF.

Source Files

Displays/hides additional options when exporting Multiple Files.

Folder Selection

Target folder for the exported TIFF files.

Include Subfolders

Even images in the subfolders are listed in the window below.

>> Feed >>

Moves the selected images from the left file list to the right. Only images in the right list will be exported to TIFF. To remove an image from this list, click .

TIFF Settings

Displays/hides additional TIFF settings.

Output Folder

Defines where the TIFF files will be saved. Same as Input can be checked to save the files into their source folder.

Folders and Naming

These combo boxes define subfolder and file naming of the newly created TIFF files.

Standard TIFF/OME-TIFF

Switches between different TIFF formats.

Store into Multipage TIFF

A multi-page TIFF file will be created containing its dimensions in one file.

Split to Multiple Files

Dimensions are dismembered to create multiple TIFF files.

Each File Contains

Defines the basic dimension(s) of the multi-page TIFF file.

Dimension of File Names

Order of the dimensions used for file naming.

Use Point Names

Names of XY multipoints present in the image will be used.

Use Channel Names

Names of the channels present in the image will be used.

Keep Original Channel Combination

If checked, channel combination of the source image is preserved.

Bit Depth

Original bit depth of the new file can be maintained, or the file can be rescaled to 8- or 16-bits.

LUTs

LUTs of the source image can be ignored (None), automatic LUTs can be applied (Apply auto LUTs) or LUTs saved with the image can be applied (Apply saved LUTs).

Save Color Channel per Pixel

Color channels can be saved per pixel to improve the TIFF file compatibility.

Mono Image for Each Channel

Each channel will be used to create a monochrome image.

RGB Image for Each Channel

Each channel will be used to create an RGB image in channel color or in grayscale.

Burn Scale

Inserts scale into the final image. The scale becomes part of the image and cannot be removed.

Burn Binary

Inserts the binary data into the final image. The binary data become part of the image and cannot be removed.

Burn Annotation

Inserts annotations into the final image. The annotations become part of the image and cannot be removed.

All Channels Merged to RGB Overlay Image

One RGB image is created from all channels available in the source image.

Note

Bit depth conversion cannot be used with spectral images.

Include OME Metadata

OME metadata can be saved with the TIFF file to increase the TIFF file compatibility (e.g. proper channel designation).

Save As

Saves the dialog window settings into the database (New...) or .xml file (Save to XML file...).

Load

Loads the dialog window settings from the database or .xml file (Load from XML file...).

Remove

Removes the selected dialog settings from the database.

Export

Starts the TIFF exporting process.

Cancel

Closes the dialog window without applying any settings.

Help

Opens this help page.

File > Save to Database

(requires: DBASE (Database))

Saves current image to database. This command displays the following dialog box:

Replace image in its original table

This option becomes active when you use the Save To Database command on the same image for the second time. If you want to replace the first copy with this second one, select it.

Save image as new record in any table

Choose the table to save the image to.

Flags

Check the image layers you wish to save within one database record. Enables you to save the binary or the color layer as a stand-alone image. The color image is saved every time.

Save

When you press Save, the Database Record Fields window appears to be filled. Fill it in and press OK. If you use the command repeatedly, the Use Last Values button can fill the most recently used values inside the Database Record Fields window.

File > Close

Closes the current image. If the image has been changed or is new, the user will be given a chance to save it.

See Closing Images.

File > Reload

This command reloads the last saved version of the current image from hard disk. If the image has been changed since the last save, a confirmation dialog appears.

File > Duplicate

This command creates a copy of the current image. Note, that the copy is created in a memory only and should be saved if you want to keep it.

File > Delete

Deletes the current image from disk. A confirmation dialog appears.

File > Import/Export > Convert Files

Converts files to different image formats. The Convert Files dialog box appears:

Directories

Specifies image file directory.

Selected Images

Lists all images of the selected type in the currently selected Directory.

File Names

This box allows you to narrow the selection of the files to convert. The Source box lists files that have the filename extension selected in the extension box below. To select a list of files that begin with particular letters, type the letters here.

File Types

Specifies filters for image files in the selected Directory.

Output Directory

Destination directory path. Press the Same as Input button to set quickly the same directory. If you want to choose a different output location, press the ... button and pick up another directory.

Output Size Parameters
Keep Original Size

Maintains image's width and height.

Set Custom Size

Changes image's width and height to specified values.

Keep Aspect Ratio

If checked, height of the image is automatically recalculated to preserve aspect ratio.

Scale down the maximum dimension

Scales down the image. The longer side of the image is rescaled to the defined number of pixels while the aspect ratio is kept. Shorter side of the image is calculated automatically.

Output File Info

This option is available only for the JPEG2000 and the LIM format. You can keep original info or you can set a new one.

Keep Original Info

Keeps the original file info setting.

Set Custom Info

Allows to set custom image fields values. To set the new info, press the File Info button.

Include Subdirectories

If checked, images in all subdirectories of the specified directory will be converted too.

Overwrite Original Files

Check this item to overwrite the original images with the converted ones.

Output File Format

Specifies destination image file format.

Compression

Specifies type of compression of the destination image.

OK

Starts the file conversion and opens the Conversion Progress dialog box. The dialog box displays messages about the file conversion's progress. To stop the conversion, click the Abort button.

File > Import/Export > Crop ND Document

Any ND2 file can be cropped. You can see the four dimensional image window in the example picture.

To crop the ND2 file
Adjust the Image Selection

A red rectangle surrounding the current image appears when the Crop ND command is called. Move the window aside and adjust the rectangle size and position by mouse. The position and dimensions of the selection are indicated in the window. Everything that exceeds the red border will be cropped.

Restrict the Time Dimension

The white bar in the Time Dimension section indicates single frames. Move the green sliders to determine which frames are to be cropped. You can also fill the range in the below edit boxes. When the step is set for example to โ€œ2โ€, every second frame will be included in the resulting ND2 file

Restrict the Z Series Dimension

This dimension can be adjusted equally to the Time dimension. The Z low and Z high values of the green sliders are indicated in the window.

Restrict the Wavelength Dimension

This is done by unchecking the channel that you would like to discard.

File > Import/Export > Split Multipoints

When there is an ND2 file that includes more than one XY - multipoint position, this command enables you to separate these points. This results in a number of non-XY ND2 files. The following window appears:

  • Select the destination directory where the files will be stored.

  • Define the prefix to determine the resulting file names. A number will be appended to this prefix.

  • Select the Split button.

  • The window closes and the files will be created in the destination directory (they will not open automatically).

Note

If well names are stored inside the ND document, they are automatically used for naming of the resulting files (e.g. A1, A2, โ€ฆ instead of 1, 2, โ€ฆ).

File > Import/Export > Create ND File from File Sequence

An ND2 file can be composed from a sequence of images. Its structure can be defined via the following window:

ND Experiment Shape
Get from File Names

Check this option if image names contain information about the the dimensions. Eg.: seq_Z01T01.jpg, seq_Z01T02.jpg, etc.

Define Manually

If the image sequence is not indexed properly, check this option

Set the ND Experiment Shape to the Define Manually mode.

  1. Select the dimension types from the combo boxes to define the dimensions order.

  2. Define the number of loops for each dimension in the Statistics portion of the window.

  3. Set the filtering conditions through the Advanced File Filter dialog.

  4. Select one file of the files in the list to determine the image sequence (images with the same prefix are automatically selected).

  5. Specify the Dimensions properties in the right portion of the window.

  6. Confirm the options by the Convert button.

Custom

Check this box to enable custom filename filtering. You can use these wildcard characters: โ€œ#โ€ matches any digit and โ€œ?โ€ matches any character.

Time, Multipoint, Z Series, Wavelength, None

From the four pull-down menus, select dimensions to be created. Set the dimensions of ND2 file (Time, Z Series, Wavelength, Multipoint). Specify number of dimension frames in file sequence in the field below. This option is enabled for Define Manually option.

File list view

This portion of the window displays a list of all available files. Select the file which will represent a first frame in ND2 file by double-clicking its name or click on the point before the file name.

Timelapse

Set Timelapse period value. Specify the exact value or select to use File Creation Time or Acquisition Time.

Z Series

Set the Z Series step as a value (ยตm) or select to use the Z Series step from Image Data.

Wavelength

Define name of the channel. Define the optical configuration name. Choose one of the seven predefined colors or brightfield option as a color of the channel. You can also select the [More] option to define a custom channel color.

Open the file afterwards

If checked then the created ND2 file is opened in the application after the conversion.

Convert

Create the ND2 file according to the given properties.

How To

Create a Common ND2 File Automatically

This is the easiest way of creating the ND2 file. All you need to have is a properly indexed source data, which typically means to have sequence of images named like this:

prefix index1 string index2 string index3 .. e.g.: โ€œseq_Z1T1โ€.

string , prefix - is recognized as any combination of letters. โ€œseq_Zโ€ stands for a prefix and โ€œTโ€ for a string in the example.

Note

It is possible to use the filenames in the form [*010101.*] with [Create ND File from File Sequence] dialog to automatically create ND file with the dimension coordinates specified by the filenames.

Set the ND Experiment Shape to the Get from File Names mode.

  1. Browse for the image sequence within the Directories portion of the window.

  2. All images of the selected directory appear in the list of files in the middle of the window.

  3. There might be several sequences in the directory. Just click on one file of the desired sequence and it will be detected automatically (under condition of having different prefixes). Other files will look disabled (gray).

  4. When a sequence is selected, the filenames are automatically searched for the indexes, and the indexes are marked with color according to dimension types selected in the above pull-down menus.

  5. Change the dimension order by selecting from the pull-down menus, if needed.

  6. Check, whether the system recognized the right image order, and define the properties of selected dimensions in the rightmost portion of the window.

  7. Click Convert to start the ND2 file creation. The file will be placed as defined in the Output File Name field.

Dimensions properties

The Timelapse properties enables you to set the time between images manually (specify period), or to count the time from file attributes (Read File Creation time), or use the records stored inside the files captured with NIS-Elements (Read from Acquisition Time).

The Z-Series properties requires to define a z-step. This can be filled in manually or let the system read it from the image data.

You can define the Wavelength properties: the name, the color, and the optical configuration that was used for capturing each channel.

Create a Common ND2 File Manually

If there is an image sequence which is not indexed properly, the ND2 file can be created manually.

Set the ND Experiment Shape to the Define Manually mode.

  • Select the dimension types from the combo boxes to define the dimensions order.

  • Define the number of loops for each dimension in the Statistics portion of the window.

  • Set the filtering conditions through the Advanced File Filter dialog.

  • Select one file of the files in the list to determine the image sequence (images with the same prefix are automatically selected).

  • Specify the Dimensions properties in the right portion of the window.

  • Confirm the options by the Convert button.

File > Import/Export > Create JSON File Sequence Description

Saves information about a sequence of images to a JSON file. This file can then be processed as an ND2 file without actually creating one. This is useful when conversion to ND2 would copy lots of image data (hundreds of GB), and especially when the file is to be processed further more in NIS-Elements (e.g. Deconvolution, Denoise, ...) which involves saving. Therefore the JSON file format is a transient format with limited functionality.

The options of the dialog correspond to the File > Import/Export > Create ND File from File Sequence command.

File > Import/Export > Export ND Document

All frames of an ND2 file can be saved as single image files by this command. Tiff and JPEG2000 file formats are supported. This window appears:

File folder

Browse for and select a folder to save a file into.

File type

Specify the new exported file type.

File prefix

Enter a text to to be used at the beginning of the file names.

Index order

The order of letters corresponding to dimensions included in the ND2 file can be adjusted. (c stands for Channels)

Apply LUTs

This option modifiest the exported image data by applying the current LUTs settings.

Insert Overlays

This option merges all visible layers (binary, annotations, ...) and inserts it to the underlying image data. If this option is used, the images are always exported as RGB images.

Channels

There are three available ways to save the ND2 file channels:

Mono image for each channel

Each channel is exported as a mono image and indexed โ€œc1, c2, ...โ€.

Color image for each channel

Each channel is exported as an RGB image and indexed โ€œc1, c2, ...โ€.

Keep image as multichannel

This option keeps the channels layout of the original ND2 file.

Saving TIF files

When saving a tif file, you can choose to keep the original bit depth of the new file, or to rescale it.

How to export an ND2 file

  1. Define the File Folder where the images will be saved.

  2. Select between the Tagged Image Format and JPEG2000 file formats.

  3. Fill in the file name prefix after which the indexes of dimensions will be placed.

  4. Select the method the channels will be treated with (see below) in the Channels section.

  5. Select the Index order. (Lets say you have two dimensional ND2 file with โ€œtโ€ and โ€œzโ€. If a โ€œt,zโ€ order was selected, the image frames would be ordered โ€œt1z1, t1z2, t2z1, ...โ€, otherwise, the order would be โ€œz1t1, z1t2, z2t1,...โ€

  6. Press the Export button.

Tip

To reduce the size of an EDF file which usually contains a Z map and other information, the user can use this command with Apply LUTs and Keep image as Multichannel on.

File > Import/Export > ND Export to TIFF

Exporting the ND document to the TIFF file. Enables the user to set the Output, Index order, TIF Compatibility Options and Channels.

Dialog window options are similar to File > Import/Export > Export ND Document.

Keep image as multichannel

This option can save the color channel data per plane or per pixel. If you are using a 3rd party software which cannot open your Per pixel exported TIFF files, choose the Per plane option and export them again.

File > Import/Export > Export Multiple ND Files

Similarly to the File > Import/Export > Export ND Document command, you can save more ND files at a time as single image files by this command. Following window appears:

File folder

Select a folder to save a file into.

File type

Specify the new exported file type.

Index order

The order of letters corresponding to dimensions included in the ND2 file can be adjusted.

Channels

There are three available ways to save the ND2 file channels:

Mono image for each channel

Each channel is exported as a mono image and indexed โ€œc1, c2, ...โ€.

Color image for each channel

Each channel is exported as an RGB image and indexed โ€œc1, c2, ...โ€.

Keep image as multichannel

This option keeps the channels layout of the original ND2 file.

TIF Compatibility Options

When saving a tif file, you can choose to keep the original bit depth of the new file, or to rescale it.

Export each ND2 info individual subfolder (named according to ND2 name)

Exports the images of each ND2 file into individual subfolders.

Export all TIFFs into one folder

Exports all the TIFF images of all the ND2 files into one folder, that you can specify, or you can use the Same as input option, to define the path to the folder.

File > Import/Export > Create ND File from BioStation NEX

This command imports a NEX file and converts it to an ND2 file.

File > Import/Export > TWAIN

Acquires images from TWAIN compliant devices (scanners, digital cameras etc...). If you have more than one TWAIN compliant device installed, the system prompts you to select the required one. Before you actually acquire the image, usually, you can adjust some scanning properties in the scan setup window. For details, please see the manual of your TWAIN compliant device. After the Acquire command has been performed, the image named Acquired 01 opens. To save it, select File > Save As. If you have acquired a series of several images without saving, the file names are numbered (Acquired 01, Acquired 02, ...).

Note

This command is available only in the 32-bit NIS-Elements application.

File > Import/Export > Select Source

Selects a source for acquiring via the Twain driver interface. Select the device, you want to use and press Select. See also: File > Import/Export > TWAIN.

Note

This command is available only in the 32-bit NIS-Elements application.

File > Open/Save Next > Save Next

This command saves the current image (live or static) automatically according to the defined settings. The destination path, file format, file name prefix and other options can be set via the File > Open/Save Next > Save Next Options command. Click the Save Next command and the following window appears. Press the OK button to save the current image.

Destination path

The destination path and the name of the image that is about to be saved is displayed in the window.

Suppress this message next time

If you check this option, next time you invoke the Save Next command, the window will not appear and the image will be saved right away.

Change...

This button calls the Save Next Options window where all the settings can be adjusted. See File > Open/Save Next > Save Next Options for further information.

Grabbing window 
If instead of the Save Next window the grabbing window appears, it means the Show grabbing dialog before saving option is selected in the Save Next Options window (File > Open/Save Next > Save Next Options). This window enables you to define the way of capturing of the current live camera signal. Set the averaging, integration, bit depth, and shading correction and then press Capture.

File > Open/Save Next > Undo Last Save Next

This command is enabled only if the File > Open/Save Next > Save Next command was used for automatic saving of images. If an image was accidentally saved (by pressing Enter), and you wish to delete it, use this command.

File > Open/Save Next > Save Next Options

This window enables you to configure the File > Open/Save Next > Save Next command properties.

Save to File
Directory, Prefix, Digits, File Format, Compression

Defines name of the file that will be automatically generated. Lets assume, that you have selected - Directory: c:\images; Prefix: seq; Digits: 4; File Format: JPEG2000. Then, by calling the File > Open/Save Next > Save Next command, an automatic file named โ€œseq0001.jp2โ€ is generated and saved to the "c:\images" directory. When pressing Enter again, the file โ€œseq0002.jp2โ€ is saved to the same directory etc.

Define Image Info

Displays a dialog box, where you can enter some description (jp2 meta-information), that will be saved with every image:

Overwrite/Skip already existing files

Allow or deny rewriting of the images already existing in the default save directory.

Save to Database

(requires: DBASE (Database))

When selected, images are being saved to a chosen database table, instead of saving them directly onto disk.

Database

If you are already connected to some database, select it from the pull-down menu. Else a dialog window appears, which enables you to connect to any database.

Table

Select the database table which the images will be stored to.

Autoincremental field

Choose which field of the database will be used to store the generated image descriptions.

The image descriptions are generated according to the Prefix, Digits, Number values. Lets assume, that you define - Prefix: seq; Digits: 4; Number: 20. Then, by pressing Enter , the current image is saved to the selected database table and the automatic image description is put into the Autoincremental field of the table. Its name would be โ€œseq0020โ€ in our case. When pressing Enter again, the second record signed โ€œseq0021โ€ is created.

External mapping

You may create an external *.txt or *.ini file containing mappings that assign table field names to some particular values. These values are automatically filled into the database records when the Save Next command is called. Please, see the comments below for an example.

Note

External mapping example - The *.ini file content may be:

[Table 1]
Author=Jack Sparrow
Experiment number=12
Sample=Malus sylvestris

It means: When saving images to the database table named โ€œTable 1โ€, the fields of Author, Experiment number, and Sample will be filled with the specified values.

Options
Show grabbing dialog before save

Before an image is saved the Grabbing dialog box is shown. Selecting this option enables you to define the way of grabbing (with/without a shading correction, averaged etc).

When saving, display Image Info dialog box

You can change the default image information (jp2 meta-information) right before saving.

Sound Alert

If checked, a short tone from your PC speaker is played every time the Save Next command is used.

Save with annotation and binary layers

If checked, images are saved together with the binary and annotation layer.

Change to Live after saving an image

After saving a single image, a Live image is displayed immediately.

Save OME Metadata

OME metadata can be saved with the TIFF file to increase the TIFF file compatibility (e.g. proper channel designation).

See also Supported File Formats.

Defaults for This Page

Pressing this button restores the default settings of this window. All your changes will be lost.

OK

Confirms all the changes and closes the window.

Cancel

Discards all the changes and closes the window.

Apply

Saves the changes which are applied, bud the window remains opened.

Help

Displays relevant help page.

File > Open/Save Next > Open Previous

The system tracks the opening and saving operations and then tries to open the 'previous' image in your working directory. Example: when you open image 'seq4.lim' by File > Open command, this command will open 'seq3.lim' image. Let's suppose that seq1.lim, ..., seq100.lim images were saved in the same directory.

File > Open/Save Next > Open Next

The system tracks the opening and saving operations and then tries to open the 'next' image in your working directory. The behaviour of this command differs:

The current file was opened via Organizer

When you set some sorting, filtering, or grouping in Organizer (for both DB records and files), and then open an image by Organizer, the applied sorting is maintained also for the OpenNext command. Furthermore, the sorting criterion is copied to the File > Open/Save Next > Open Options.

The current file was opened via the Open command

When you opened an image by the File > Open command (does not matter if in the Organizer or Image layout), the OpenNext command uses settings of the File > Open/Save Next > Open Options. It does not influence Organizer when these settings are changed.

File > Open/Save Next > Open First

The system tracks the opening and saving operations and then tries to open the 'first' image in your working directory. Example: when you open image 'seq4.lim' by Image File > Open command, this command will open 'seq1.lim' image. Let's suppose that seq1.lim, ..., seq100.lim images were saved in the directory.

File > Open/Save Next > Open Last

The system tracks the opening and saving operations and then tries to open the 'last' image in your working directory. Example: when you open the image 'seq4.lim' by Image File > Open command, this command will open seq100.lim image. Let's suppose that seq1.lim, ..., seq100.lim images were saved in the directory.

File > Open/Save Next > Open Options

This window enables you to configure the File > Open/Save Next > Open Next command properties.

Open from file
Directory

Specifies the directory with files to open.

Files of Type

Filters the files to open by the image format.

Order By

This pull-down menu enables you to select one of the image properties as the ordering criterion. The button toggles the alphabetical order.

Prefix

Filters the files to open by prefix. You can click to show advanced filtering options window. The button turns the advanced filter on/off.

Next file

Displays the name of the file that will be opened after calling the function next time. You can see its name on the right side of the box.

Open from Database

(requires: DBASE (Database))

At least one database must be connected in order to enable this option. Select a database from the list. A database can be created by the Database > New Database command.

The other options are analogical to the ones described above.

Defaults for this Page

Pressing this button restores the default settings of this window. All your changes will be lost.

OK

Confirms all the changes and closes the window.

Apply

Saves the changes which are applied, bud the window remains opened.

Cancel

Discards all the changes and closes the window.

Help

Displays relevant help page.

File > Merge Channels

Merges images displaying the same scene into one image. Each of them is usually captured under different illumination and using different optical filters and different camera settings. It combines color planes stored in separate files into one RGB image. The Merge Image Planes dialog box appears:

The function can take up to three monochrome (fluorescence) images and one color/monochrome (bright field) image. If a color image is inserted into one of the mono channels instead of the monochrome one, the appropriate image component is extracted. The bright field can be either color or monochrome. Monochrome images are transferred into the appropriate color components of the result image which is then blended with the bright field using a special formula. Each source image can be skipped if the user selects the <None> option. If bright field is skipped, the source components can be later easily extracted from the blended image by the Image > Convert > Extract Component command. If the source images are of different sizes, all of them are stretched to the maximum size - images acquired with a different camera binning or resolution can be merged.

Red/Green/Blue Component

Selects a source image (must be opened in NIS-Elements) for defined component.

Brightfield

Selects a source image for bright field.

File > Stitch Large Image from Files

Creates large images from a sequence of files that were captured separately. The following window appears:

To load sequence of images:
  1. Click Open Image Sequence.

  2. The Load Image Sequence dialog appears:

  3. Browse for the directory where the sequence is saved.

  4. Select the files to be loaded by defining the File Names restriction. (e.g. โ€œLargeโ€ will restrict the selection only to files beginning with the word โ€œLargeโ€).

  5. Click OK. The dialog closes and the selected files are loaded to the main window.

Creating Large Image
  1. The preview of loaded images is arranged inside the window according to the Columns/Rows value - depends on your choice of the Vertical/Horizontal direction. Change it to change the arrangement.

  2. Select whether the images were captured in meander or rows direction and set the position of the First frame.

  3. Choose the Overlap rate from the combo box. Select the best value according to the preview image. Auto scale automatically sets LUTs for better visualization.

  4. If you check the Image Registration option, the images will be merged using a special algorithm which tries to fit and smooth the edges.

  5. The Crop a border option crops the large image so that no background (caused by the image registration algorithm) will be included in the result. When image registration is used and the result is not cropped, it contains asymmetric black borders.

  6. Stitching option is used to select a stitching method. Blending method blends the overlapping image parts and Optimal path method computes a contour in places where two overlapping images are least different and then they are stitched copying this contour.

  7. Shading correction function is appropriate for sufficient stitching of homogeneous images. Check Automatic shading correction to turn the function on and choose the type of correction which best represents the background of your sample image. If Automatically per channel is selected, the shading modality method is detected and applied per channel. For more information about shading correction, please see: Acquire > Shading Correction Panel.

  8. When done with setting all of the options, click Stitch. When the stitching process finishes, the large image is opened as a new image called โ€œLarge Image #โ€.

File > Image Properties

This command opens the Image Properties dialog. The dialog appearance may vary according to the type of image. In case you are inspecting an ND2 file properties all the following tabs are accessible.

Image Attributes

Displays the file properties which are provided by the system. Among these properties you can find the file name and location, dimensions of the image, file size, and file date. The version of NIS-Elements including the build number in which the image has been saved is also stored in the image metadata.

Original, Modified

The software can determine whether the current image was modified after acquisition. To indicate it, a green text โ€œOriginalโ€ or a red text โ€œModifiedโ€ appears in this dialog.

The โ€œModifiedโ€ text has a different meaning depending on its placement in the dialog:

Image Attributes

General changes of image attributes like size, bit depth, number of frames.

Image Fields tab, Custom Metadata tab, Acquisition Details tab

The metadata mentioned on this tab have been changed.

Recorded Data tab

Click the Authenticate Frames button to update status of all image frames. Changes in raw image data will be detected for each frame.

Note

Image authentication can be turned off via the Edit > Options command.

Copy Full File Name to Clipboard

Full path to the current image file is copied to clipboard.

Export All Available Information

This button exports the data of all tabs at once. Select the destination where to export the data. See also Exporting Results.

Image Fields

This tab appears for all image types. These fields have changeable content. The fields are related to better file recognition, so you can for example specify the image ID, enter any description you like, write you own conclusion and many more. The line on the right side of the fields indicates relevance of the fields to file formats. LIM files use only the upper fields. JP2, tif and ND2 files contain all the fields.

Fill by Defaults

Default values can be predefined by the user (they will be prefilled every time a new image is being saved). Insert these values in the current table by the Fill by Defaults button.

Edit Defaults

Press this button to open a Default window. Specify all the default content. You can edit all fields. The content can be relative - every time you click the Fill by Defaults button, the current image data will be used, or you can enter fixed content.

, Substitutional keywords

The system enables the user to insert some predefined keywords inside the Image Information table. When saving an image, the keywords are dynamically replaced according to the current system state. E.g. the current objective name can be inserted:

The Description and Capturing fields may contain many rows of text. Use this button to view/edit it in a separate resizable window.

Experiment Data

This tab appears when you are working with ND2 files. The experiment data tab displays the information about the performed experiment. Provides a summary information about the ND experiment process: if the ND2 file contains spectral, time or z-stack loops, settings of all available dimensions are included.

Z-Step

The calibration of the Z axis may be modified.

  1. Click this button.

  2. A dialog box appears where the new Z-step can be set:

  3. Set the step size and confirm it by OK.

T-Step

If the ND2 file contains only one phase, the interval between the time loops may be modified.

  1. Click the Edit T-Step button.

  2. A dialog box appears where the new time interval can be set.

  3. Set the interval length and confirm it by OK.

New step value affects the Time Loop parameters.

Reuse

Reuse the current ND image setting for creating a new ND2 file. The ND Acquisition window appears with the experiment defined. You can start the new ND acquisition experiment by pressing the Run Now button.

Export

This button exports the data to clipboard, MS Excel application, or an external *.txt file. Select the export output from the pull-down menu next to the button.

Recorded Data

The Recorded Data tab contains a table with information about single frames ordered subsequently:

Use Zero Based Time Scale

Sets acquisition time of the first frame to 0 and adjusts all other time stamps relatively to their time offset from the first frame.

Time

Time when the frame was captured in sec. It is counted from the experiment start.

delta Time

Time difference between adjacent frames.

Index

Index number of the frame.

Z series

Z position of the frame.

X Coord, Y Coord, Z Coord, Zdrive, etc.

Other device data selected for recording during ND acquisition.

Events

Displays performed user events during acquisition.

Press the ... button to open the Edit Events window.

Select type and time of an event and append an arbitrary description.

Authenticate Frames

Performs authentication of all image frames. Frames are tested whether they have been modified after acquisition or not. The Enable authentication option must be turned ON in Edit > Options , otherwise this button is hidden.

Export

This button exports the data to clipboard, MS Excel application, or an external *.txt file. Select the export output from the pull down-menu next to the button.

Custom Metadata

Any custom-defined metadata can be stored inside ND2 documents. This metadata can be defined in Image Properties, ND Acquisition and JOBS.

Add/Modify Description

Opens the Modify Description dialog window where metadata can be altered.

Export

This button exports the metadata to MS Excel, external *.txt file or to clipboard. Select the export output from the pull-down menu next to the button.

Acquisition Details

This tab displays information about the hardware settings used to capture the image. The following values can be displayed:

Camera name
Optical configuration name
Microscope name
Emission and Excitation wavelengths of each channel
Channel modality of each channel
Refraction index and numerical aperture of the objective used

Expand Channels, Collapse Channels

Use these buttons to expand or collapse the tree with channel descriptions.

Enable Editing

Click the button to unlock the right column where you can edit information about the channels. Select new values from pull-down menus (Channel modality) or double-click the fields to edit them (emission wavelength, excitation wavelength, ...).

Export

Press this button to save the acquisition details to a .txt file. A standard Save As window appears.

File > Print

Prints the current image. The Print dialog box appears.

Print Window Options

Printer

Shows the currently selected printer.

Source Rectangle

Specify the portion of the image you want to print.

All

Prints the entire image.

Selection

Specifies a rectangle within the current image only which will be printed.

Destination Rectangle

Specify how the image will be arranged on the page.

Maximize Image

Printed image will match the maximal printable area. This depends on the printer type and actual settings.

Size

Allows you to set size of the image on the printed page.

Printout/Image Ratio

In case of calibrated images, a scale factor can be used to define the printout picture size. The value defines the ratio between the printout size and the real image size. For example, the current image dimensions are 640x480 px and the calibration is 1.5 ยตm/px. Set the Printout/Image Ratio to โ€œ100โ€ and the printout size will be (640*1.5*100)x(480*1.5*100) ยตm = 9.6x7.2 cm.

Manually

Allows you to set image width and height manually.

Keep Aspect Ratio

If checked, height of the image is automatically recalculated to preserve the height/width ratio of the original image.

Width

Specify the image width (in mm) on the printed page.

Height

Specify the image height (in mm) on the printed page.

Position

Allows you to set position of the image on the printed page. The At Center option centers the printout horizontally/vertically, the other option allows you to specify a margin in the X/Y axes directions.

Composed printing - Enable

Prints multiple images on one page. Check this option and press OK. The window will close but nothing is sent to the printer. Open another image and run File > Print again. The current image is added to the page. The print window will behave this way until you select the Print composed page option (which appears in the place where the check-box used to be).

Num. of Copies

Specifies the number of copies you want to print.

Orientation

Choose the paper orientation, โ€œlandscapeโ€ or โ€œportraitโ€.

Sublimation Printer

This option optimizes print for sublimation printers.

Setup

This button opens the printer settings window.

File > Report > New Blank Report

This command runs the Report Generator. Please see Creating Reports.

File > Report > New Report from Template > Browse

Creates a new report from the selected report template. See Report Templates.

File > Report > New Template

This command runs the Report Generator and opens an empty report template. Please see Report Templates.

File > Report > Open

This command runs the Report Generator and displays a dialog window for selecting a report file (rpt, rtt) to be opened.

File > Report > Recent Files

Opens a report selected from the list of recently used reports.

File > Report > Manage Shared Reports

This command opens a window for sharing and managing the shared reports and templates. Users with the right to Modify shared reports can import, remove, and edit any of the shared reports and templates. Users without this permission can only use them to create reports of their own.

Import

This button adds a report or a report template to the list of shared reports. A standard open-like window appears. Select the report to import to the list.

Note

The imported files are copied to NIS-Elements application data directory. The original file is not connected to the list of shared reports.

New Report from Template

If a report template (*.rtt) file is selected in the list, the button enables you to create a new report out of it. The Report Generator opens.

Edit

Selected reports/report templates can be modified within the report generator using this button. Users with the Modify shared reports right can modify them and save the changes. Other users can save the modified reports to a new file.

Remove

Users with the Modify shared reports right can remove a selected report/report template from the list.

File > Report > Send Current Image to Report

This command creates a new report, and inserts the current image to the report page. If performed repeatedly, the images are placed one after each other to a single report. Please see Creating Reports.

File > Recent Files

Opens the selected file. Up to 16 most recently opened files are remembered in the Recent Files list.

File > Recent Folders

Up to 16 most recently opened directories are remembered in the Recent Folders list. Click on one path in the list to reveal the Open Image window enabling to select and open image files from this directory.

File > Exit

This command will close NIS-Elements and also save its current configuration.