Quick Operation Guide

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Quick Operation Guide Power ON Mounting specimens Set the specimen on the sample holder, and install the sample holder to the holder frame. Attach the holder frame to the XY stage.

Slide glass Cover glass Lens For mm dishes: Set mm dishes on the sample holder ( mm dish ). For a well plate or flask: Set a multi-well plate or a flask directly on the holder frame.

CFI Plan Fluor DL 0x S PL FL ELWD ADM 0xC S PL FL ELWD ADM 0xC CFI Plan Apo λ 60xH CFI Plan Apo λ 00xH Magnification x x 0x 0x 0x 0x x 0x 0x 0x 60x 00x WD (mm) 8.0 0.00.00.0.00 0. 6.0.0 8. to 6.9.

1 Quick Operation Guide Power ON Mounting specimens Set the specimen on the sample holder, and install the sample holder to the holder frame. Attach the holder frame to the XY stage. Type of holder Main power ON Top panel OPEN For slides: Set slides on the sample holder 68.8 mm (slide ). Note: When mounting specimens, invert slides so the cover glass faces downward. Slide glass Cover glass Lens For mm dishes: Set mm dishes on the sample holder ( mm dish ). For a well plate or flask: Set a multi-well plate or a flask directly on the holder frame. For a large dish or oversized slide: Set large dishes or oversized slides on the regular versatile holder. Power ON Changing lenses and magnifications The lens currently used is displayed. Lens change Click the lens button. Basic Lenses Phase Lenses Oil Lenses BZ-X800 compatible lenses Lens name CFI Plan Apo λ x CFI Plan Apo λ x CFI Plan Apo λ 0x CFI Plan Fluor 0x CFI Plan Apo λ 0x CFI Plan Apo λ 0x CFI Plan Fluor DL x CFI Plan Fluor DL 0x S PL FL ELWD ADM 0xC S PL FL ELWD ADM 0xC CFI Plan Apo λ 60xH CFI Plan Apo λ 00xH Magnification x x 0x 0x 0x 0x x 0x 0x 0x 60x 00x WD (mm) to to Phase contrast This list shows the BZ-X800 lenses and their compatible container types. Oilimmersion Slide/Glass bottom (Thickness 0.7 mm) Plastic container (Thickness mm or more) Click the icon of the lens to adjust. Lens adjustment When applying oil or adjusting a correction ring, use the lens adjustment function. Click [Adjust Lens]. Click the icon of the lens to adjust. Open the top panel or front panel, and then apply oil or adjust the correction ring. When the adjustment is complete, click [OK].

Basic Operations Switching CCDs Switching filters Click [Mono] or [Color] to switch the mode

Image quality settings: Select [Standard] for fluorescence observation.

channel button to enable observation with the set fluorescence filter and observation method.

exposure time. Clicking [Auto] adjusts exposure automatically.

Clicking [Auto-Focus] moves to the focus position automatically.

Tip You can adjust the focus by keeping on pressing the key on the keyboard with moving the

[Ctrl] + [Shift] key + wheel [Ctrl] key + wheel [Shift] key + wheel Super rough operation

click [Focus Up] again. X-Y movement Clicking on the stage view performs the XY movement.

Black balance/white balance Black balance (Fluorescence observation) Drag the rectangle frame

White balance (Brightfield observation) Observation of fluorescence signals is much clearer

2 Basic Operations Switching CCDs Switching filters Click [Mono] or [Color] to switch the mode to monochrome observation (pseudo color) or color observation. Image quality settings: Select [Standard] for fluorescence observation. High Resolution Standard High Sensitivity 90 0 pixels pixels pixels Click a channel button to enable observation with the set fluorescence filter and observation method. Adjusting exposure (brightness) Adjusting focus Dark Bright Click the buttons to adjust the exposure time. Clicking [Auto] adjusts exposure automatically. Low photobleach mode: When observing fluorescent samples, set [Low Photobleach] to ON. Clicking [Auto-Focus] moves to the focus position automatically. To adjust the focus, right-click the mouse to select [Z Focus], and rotate the mouse wheel. Tip You can adjust the focus by keeping on pressing the key on the keyboard with moving the mouse wheel as well. [Ctrl] + [Shift] key + wheel [Ctrl] key + wheel [Shift] key + wheel Super rough operation Rough operation Slight operation Note: If the bottom surface such as the plate is in focus, click [Focus Up] again. X-Y movement Clicking on the stage view performs the XY movement. Using the well plate, the stage moves to the center of the well by double-clicking. Black balance/white balance Black balance (Fluorescence observation) Drag the rectangle frame on the background to be black on the screen, and click the set. White balance (Brightfield observation) Observation of fluorescence signals is much clearer after using Black Balance. Double-clicking on the observation screen moves the position to the center of the screen. The XY movement can be made by dragging on the screen as well. The stage moves to the pressed arrow direction. Pressing the [C] key moves to the original point. In the white balance, the degree of color is adjusted based on the area specified by.

Multi-color capture Click [Multi-Color].

3 Multi-color capture Click [Multi-Color]. Click each channel to overlay and individually adjust the brightness and focus. The overlaid image of the selected channels is displayed on the screen in real time. Click [Capture] to save the images together in a group. Note When the field-of-view is moved, click [Recapture All]. All channels are updated to display the new location. Analyzer The enhanced edit and analysis can be made by using Analyzer. Level correction Scaling The Ctrl + mouse wheel enables scaling. Adjusting the level correction can display a dark image to be brighter. Scale inserting An arbitrary length can be displayed by: Inserting Scale on the menu bar. Trimming The image trimming can be made by specifying area and cutting the image.

Advanced Functions Navigation Use this function to display the entire sample, and to quickly move to areas of interest by

Click the location to observe on the navigation screen.

The registered locations are also displayed on the navigation screen.

If the grid pattern does not display as clear, distinct lines, move the slide bar to the right.

Click [Auto] for Brightness. Saturated pixels are displayed in the set color.

4 Advanced Functions Navigation Use this function to display the entire sample, and to quickly move to areas of interest by clicking the desired location on the Navigation image. Display the starting location to register. Click [Navigation]. Click [Add] on the navigation screen. Image capture begins with the specified position in the center, and creates the navigation image. Click the location to observe on the navigation screen. Clicking [Set] for Point Memo allows up to 0 capture locations to be registered. The registered locations are also displayed on the navigation screen. Sectioning capture Select [D slit] for the type and set the slide bar at the left end (0). If the grid pattern does not display as clear, distinct lines, move the slide bar to the right. Adjust the position and focus of the image on the observation screen, and then click [Sectioning]. Click [Auto] for Brightness. Saturated pixels are displayed in the set color. Confirm that the image contains no saturated pixels prior to capture. Click [Preview] to view the sectioning image. Click [Capture] to save the image. Setting capture conditions for separate locations The Multi-Point function allows up to 999 different locations to be sequentially captured and saved using individual capture settings (including magnification, channel, observation method, exposure time, etc.). Click [Multi-Point]. The [Multi-Point] button turns blue. Select the [Use Individual Setting] check box, select the desired area for registration on the stage view, and click the [Set] button. The capture condition is displayed if clicking [Setting]. Capture locations are displayed as points on the stage view. Click [Start Capture] to automatically capture all set locations.

Image stitch (wide-area continuous capture) If the whole image cannot be

Method Setting the capture range automatically Specify the capture range by

Method Specifying the number of images Specify the capture range by setting the

Select [Set Center and Number of Images] and enter the number of images to

Note The tilt correction can be made based on the Z-axis location of the

If the influence from the tilt is big because of the shallow depth such as by

Click the [Start Capture] button to start the capture.

" Or, double-click a group capture information file (*.

Loading stitched images with Analyzer starts WideImageViewer.

5 Image stitch (wide-area continuous capture) If the whole image cannot be accommodate, the stitched image can be captured easily. Click [Stitching]. Method Setting the capture range automatically Specify the capture range by setting the outermost edges, and the number of images to be captured is calculated automatically. Select [Set Edge Points] and click [Set] on the edge of the area to capture. Method Specifying the number of images Specify the capture range by setting the central field-of-view and the total number of images. Select [Set Center and Number of Images] and enter the number of images to capture in the horizontal and vertical directions. Note The tilt correction can be made based on the Z-axis location of the specified point in the edge specification. If the influence from the tilt is big because of the shallow depth such as by high magnification lens, using the edge specification can capture images better. Click the [Start Capture] button to start the capture. If the capture completes, click "Open with the Analysis Software." Or, double-click a group capture information file (*.gci) in the folder with captured images. Loading stitched images with Analyzer starts WideImageViewer. Click [Load] Click [Start Stitching] An image of which resolution exceeds 080 x 07 cannot be handled in the analysis software, so the resolution is made small. If you want to save it with the same resolution as the original, save it as a KTF file by Wide Image Viewer. Wide Image Viewer opens The file of KTF is the original KEYENCE file format which handles a high resolution image with high speed.

Advanced Functions Z-stack This function

order to assemble a fully-focused image.

Focus the upper limit of the object, and click

Focus the lower limit as well, and click the

Click [Auto] to automatically set the Z-stack

Click [Start Capture] to save the Z-stack as a

If loading the captured image by the analysis

by extracting objects by brightness etc.

Start the hybrid cell count from Analyzer.

extracted. Adjust the extraction area accordingly.

6 Advanced Functions Z-stack This function automatically captures multiple focal planes in order to assemble a fully-focused image. Click [Z-Stack]. The [Z-Stack] button turns blue. Focus the upper limit of the object, and click the [Set] button on the upper limit. Focus the lower limit as well, and click the [Set] button on the lower limit. Click [Auto] to automatically set the Z-stack pitch. Click [Start Capture] to save the Z-stack as a group. If loading the captured image by the analysis software and perform the full focus, multiple images are composited to one image. Hybrid Cell Count This can convert into numerals by extracting objects by brightness etc. by Analyzer. Start the hybrid cell count from Analyzer. Select the type of the image, and click the [Start] button. Set a threshold value of brightness to be extracted. Adjust the extraction area accordingly. The extraction result can be output by histogram or CSV. Furthermore, BZ-X800 can utilize the image cytometer function that can output statistical data of wells by observing multiple samples simultaneously. keyence@keyence.com Copyright (c) 08 KEYENCE CORPORATION. All rights reserved M86

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