KEYANCE BZ-X800/BZ-X810

Table of Contents
Hardware…………………………………………………………………………2
Basic Operations……………………………………………………………….3-10
Starting Up………………………………………………………………3-5
Getting in Focus………………………………………………………….6-8
Navigation……………………………………………………………9-10
Advanced Operations…………………………………………………………11-20
Z-Stack………………………………………………………………….11-14
Image Stitch…………………………………………………………15-17
Sectioning…………………………………………………………….18-19
Capturing Multiple Images at Once……………………………………..20

1Hardware

1. Top Panel- for mounting specimens or changing lens
2. Front Panel- for installing or replacing filter cubes
3. Power button- turns on Keyence
4. Power indicator- illuminates when power is on
5. Transmitted light unit- the top panel houses the LED light; flip it up whenmounting a specimen
6. Stage/Sample Holder- moves sample left, right, up and down
7. Objective lens- used to view the sample
1. TopPanel

2. Front Panel

3. PowerButton

4. Power Indicator

5. Transmitted LightUnit

7. Objective Lens
6. Stage/Sample Holder

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Starting Up

8. Turn on power switch of the controller on the back if not already on
9. Press the power button on the main unit. Power indicator should be blue.
10.Log in to the computer with gator link username and password
11.Double-click the BZ-X800 viewer icon. A menu screen with various options
should pop up.

12.Click "objective lens"
13.Select the lens that are mounted in order and click ↑14.The lens should be put in the order shown:

15.After the correct objective lens are put in place, click "Filter Cube"316.Enter in the correct filter cubes in the window as they correspond to thefilter cubes installed in the microscope, as shown here:

17.If you are using a plate, select "Well Plate Settings" and select the number ofwells in your plate.
18.Under the Normal tab, click "Capture Still Images"

419. A window will pop asking what type of sample older you are using. Choose
the correct one.
Sample holder

Slide holder (goes on sample holder)

Well plate holder
35 mm dish holder (goeson sample holder)

Large dish holder

20.Choose the correct holder for your sample. Open the top panel and lift thetransmitted light unit.
21.Place the sample holder over the objective lens. If placing one of theholders, then they need to be screwed and unscrewed into the stage.

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Getting in Focus

22.With the top panel still open, take note of where the sample is, thennavigate to the stage view and use the control panel or click on the slides toview your sample under the objective lens. Once the sample is in view on
the screen then this step is complete.

23.Under the "Lens' tab in the top right corner, click on the objective lens andselect the lowest magnification lens6

24.Under the "Z-position" tab, select "auto-focus". You can also manually focus
the specimen using the mouse scroll as course adjustments and ctrl+mousewheel is for fine adjustments.

25.Repeat steps 2-3 with each desired objective lens starting from lowest
magnification to highest.
Note-The higher the magnification is, the more difficult it will be to
get the image in focus. For oil immersion lens (60x), please makesure to use a drop of oil on the lens. If image is too difficult to getin focus, try adjusting the exposure and white balance.
26.Click "Mono" or "Color" to switch the mode from monochrome observationor color observation.

27.Click the "White Balance" option. The degree of color is adjusted on the area
specified by the square. This operation is telling the software what is whiteand what is not and clears out any noise in the image.

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28.Click a channel button to view a fluorescence filter. If not viewing andfluorescent samples, stay in CH4.

29.If observing fluorescent samples, click "Low Photobleach" to ON. This modeis designed to pulse excitation light only at the time of image capture so thatthe sample is not exposed to light for prolonged periods of time.

8Navigation

30.Click on the tab on the top bar menu called "Navigation"

31.Follow the steps in the pop-up window.
32.When clicking the "add" button, wait until the entire sample has appearedon the window and then hit stop.
33.Turn the objective lens to a higher magnification and get the image intofocus

934.Under the Camera Area Settings, set points can be placed on the image formultiple capture points.

35.Then click "Start Capture" at the bottom
36.Save the images into desired folder.

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Z-stack

37.Click "Z-stack" on the control panel

38.With image in focus, using the mouse wheel, scroll up just as the image
becomes blurry.

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39.Next to the upper limit, click "Set"

40.To set the lower limit, scroll down just as the image becomes blurry and click"Set"
41.Change the pitch to the recommended pitch values that correspond to theobjective lens that is currently on the image:
2x objective lens------60 micrometer
4x objective lens------20 micrometer10x objective lens-------5 micrometer
20x objective lens -------3 micrometer
40x objective lens-------2 micrometer
60x objective lens--------1 micrometer

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42.Click "Start Capture"

43.Chose a file to save the images, after images have been taken, click "AnalyzeImages"
44.Once the analyzer mode pops up, click the "load" button at the bottom of
the window.

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45.In the analyzer window, click "Full Focus"--> "Auto"-->"Save"

Note- this mode allows users to view one image captured at different focal
points to create a precise photo.

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Image Stitch

46.Click "Stitching" on the control panel.

47.Method 1- Click "Set Edge Points". Set points around the sample in order toview the entire sample under a certain magnification. (This is easier toperform when Navigation mode is on.)
48.Method 2- Click "Set Center and Number of Images". Increase the squaresize to increase the field of view or the image.

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49.Click "Start Capture" at the bottom of the screen.

50.Select a file to save the images, after the images have been captured, click"Analyze Images"--> "Load" in the Analyzer Mode.

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51.Once in the next window, under the "Image Processing" tab, click "Image
Stitch"--->"Load a Group"--->"Load"
52.A new window should pop up. Click "Start Stitching"

53.Save the image into a folder

17Sectioning

54.With the image in focus, click "Sectioning" in the control panel.

55.Chose the desired fluorescent filter.
56.Select "1D slit" and set the slide bar to the right until clear, distinct lines areseen.

1857.Click "Auto" for brightness
58.Click capture to save the image
Note- This mode illuminates the sample using a grid projecting pattern. With this,high resolution images can be captured.

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Capturing Multiple Images at Once
59.Click "Multi-Point" on the control panel

60.Next to the "Area" tab, select the area you want to capture, then click "set".
(This is best used with navigation mode in use.)
61.After all the points are set, click "Start Capture".

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