all-in-one fluorescence microscope – no darkroom …

CELL COUNTING ISSUE

All-in-One Fluorescence Microscope BZ-X800E

ALL-IN-ONE FLUORESCENCE MICROSCOPE – NO DARKROOM REQUIRED

CONCEPT·FEATURES·APPLICATIONS

5VOL.

An easier approach to fluorescence microscopy

Advanced functionality without compromising simplicity

Image Quality:Observe high-resolution optical slices comparable to a confocal laser microscope.

Application Diversity:Combine the functionality of a slide scanner and time-lapse incubator on a single platform.

Analysis Capability:High-throughput quantification software for high-content data acquisition.

Install the microscope in a location that provides the

highest experiment efficiency.

All operations are performed in the software. Any user can easily capture images and reproduce

results.

High-sensitivity monochrome cooled CCD camera provides high resolution images

in fluorescence, brightfield, and phase contrast.

Built-in Darkroom Clear ImagesFull Electronic

Controls

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P.4 CARDIAC FIBROSISMeasuring the cardiac fibrosis ratio of a crab-eating macaque

COLONYP.5 Counting iPS cell colonies in a well plate

P.6 FAT CELLQuantitative analysis of adipocytes

P.8 STOMACH CANCERQuantification of immunostained stomach cancer tumor sections

P.7 FISHCounting chromosomes in a nucleus

P.9 CULTIVATED CELLSMeasuring the transfection efficiency of cultivated cells

TA B L E O F C O N T E N T S

All-in-One Fluorescence MicroscopeBZ-X800E

NEW

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BZ-X800E APPLICATION EXAMPLES

FIBROSISMeasuring the cardiac fibrosis ratio of a crab-eating macaque

Using the BZ-X800E

For a large section that cannot be seen within a single field-of-view, the motorized XY stage can capture and stitch together the entire region of interest in high resolution.

Fibrosis is a condition where tissues stiffen due to the deposition of collagen fibers.

As fibrosis progresses, normal organ function deteriorates.

IMAGE STITCHINGUp to 50,000 × 50,000 pixels can be joined together easily to capture a clear image

without stitch lines or brightness variations. Additionally, it is possible to accurately

quantify information such as fluorescent signal intensity.

Objective lens : CFI Plan Apo λ 4×Image stitching : 20 images × 12 images

AREA (TOTAL) AREA (FIBROTIC AREA) AREA PERCENTAGE

577,439,588 μm2 47,847,466 μm2 8.3%

COUNTEven for tilted specimens or specimens that have height differences, it is possible to create an image in which the entire specimen is in focus. This is accomplished by capturing multiple images in the Z-direction and stacking together only the parts that are in focus.

Hybrid Cell Count software can extract only fibrotic areas from the entire section and calculate the proportion automatically.

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Objective lens:CFI Plan Apo λ 10× Image stitching: 7 images × 9 images

Batch measurement under the same conditions

COUNT 181

AVERAGE AREA 5,133 μm2

STANDARD AREA DEVIATION 8,730 μm2

TOTAL AREA 929,027 μm2

COUNT 251



TOTAL AREA 2,086,967 μm2

COUNT 123




COUNT 171




COLONYCounting iPS cell colonies in a well plate

Induced pluripotent stem cells (iPS cells) are generated through the introduction of

specific genes into somatic cells. After cultivating for, they change into pluripotent

stem cells that have the ability to differentiate into cells of various tissues or organs and

proliferate almost without limit.

Using the BZ-X800E

For a well plate that cannot be seen within a single field-of-view,the motorized XY stage can capture and stitch together the entire region of interest in high resolution.

Hybrid Cell Count software allows extraction of only colonies from a well plate hole for automatic counting.

Multiple images can be processed collectively under the same conditions based on the features extracted with Hybrid Cell Count.

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COUNT AVERAGE AREA STANDARD AREA DEVIATION

1485 169 μm2 403 μm2

Objective lens: CFI Plan Apo λ 20×

Batch measurement under the same conditions

COUNT

COUNT

1955

AVERAGE AREA

122 μm2

STANDARD AREA

DEVIATION

352 μm2

FAT CELLQuantitative analysis of adipocytes

Adipocytes are composed of droplets of fat called adipose tissue, and can be

classified into two types - unilocular (white) adipocytes and multilocular (brown)

adipocytes. A white adipocyte contains a single large fat droplet that stores fat.

A brown adipocyte contains multiple smaller droplets that burn fat to generate

heat. If white adipocytes grow in size due to an increase in obesity, they become

dysfunctional, and can lead to the onset of certain diseases.

Using the BZ-X800E

Each closely-spaced adipocyte can be extracted and counted using Hybrid Cell Count measurement software.

The image can be filtered by the area or shape of the extracted cells.

Multiple images can be processed collectively under the same conditions based on the features extracted with Hybrid Cell Count.

COUNT

3162

AVERAGE AREA

68 μm2

STANDARD AREA

DEVIATION

186 μm2

COUNT

1190

AVERAGE AREA

222 μm2

STANDARD AREA

DEVIATION

416 μm2

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Signal G\R 0 1 2 3 4

0 40 5 1 1 0

1 23 35 13 1 0

2 10 24 18 2 2

3 0 5 7 2 0

4 1 3 4 2 1

Normal observation

COUNT

SECTIONING

No. of nuclei containing respective signals

Objective lens: CFI Plan Apo λ 100 × HSectioning + Z-stack

FISHCounting chromosomes in a nucleus

FISH (Fluorescence In Situ Hybridization) is a method of detecting the

distribution or amount of specific nucleic acids (DNA or RNA) using

fluorochromes. It is mainly used for detecting chromosomal abnormalities.

Using the BZ-X800E

The Optical Sectioning function makes it possible to eliminate fluorescence blurring and capture clear images.

A built-in Z-stack function captures multiple images at different focal positions and is able to create a fully-focused image by combining only the areas that are at their sharpest focus.

Hybrid Cell Count can be used to specify nuclei as mask areas to extract chromosomes contained in each nucleus and count them.

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Objective lens: CFI Plan Apo λ 10×Image stitching: 20 images × 12 images

AREA (TOTAL) 65,426,415 μm2

AREA (TUMOR) 10,085,121 μm2

AREA PERCENTAGE 15.4%

COUNT

STOMACH CANCERQuantification of immunostained stomach cancer tumor sectionsImmunostaining, which is also called an immune antibody method or immunohistochemistry, is

often used for histopathological diagnosis. It binds a specific protein (antigen) to tissue with a

substance that specifically reacts to it (antibody) in order to visualize tumors or other target parts.

Using the BZ-X800E

For a large section that cannot be seen within a single field-of-view, the motorized XY stage can capture and stitch together the entire region of interest in high resolution.

Even for tilted specimens or specimens that have height differences, it is possible to create an image in which the entire specimen is in focus. This is accomplished by capturing multiple images in the Z-direction and stacking together only the parts that are in focus.

Hybrid Cell Count can extract only tumoral areas from the entire section and calculate the proportion automatically.

8


Counting cells using a phase contrast image

Counting expression events (fluorescent points) using cells as mask areas

Objective lens: CFI Plan Fluor DL 10× Cell 903

Expression 89

Efficiency 9.9%

CULTIVATED CELLSMeasurement of the transfection efficiency of cultivated cells

Transfection is a process of introducing nucleic acids into animal cells.

It is used to make cells introduce specific genes to express a protein of interest.

Using the BZ-X800E

It is possible to obtain phase contrast and fluorescence overlay images.

Even from a phase contrast image, our original measurement algorithm allows the outlines of cells to be extracted accurately.

Hybrid Cell Count can be used to extract the fluorescent protein contained in the cells and count them by using cells as mask areas.

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HYBRID CELL COUNTHigh-accuracy quantification

Mask functionBrightness (intensity) measurement

Extract only the sections with a brightness within a specific range to count the number or measure the area. Brightness variation correction and other functions ensure quantification of desired targets.

Color (hue) measurement

Extract only the sections with a color within a specific range to count the number or measure the area. Detailed and precise quantification is possible even with images with low contrast or clusters of cells.

Using the first specified area as a mask allows for example, counting the number of fluorescence-expressing cells or measuring the area of malignant tumor cells in normal tissue.

Phase contrast image (outline) measurement

The outlines of cells are accurately extracted even when the image has low contrast between the mea-surement target and background, as it the case with phase contrast images. Even adjacent cells can be counted accurately and easily.

Measurement results can be output in spreadsheet format as well as be displayed by the application software. This allows flexible data editing such as graph creation according to specific purposes.

Mask creation Measurement inside mask areas

Data can be output in spreadsheet format

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MACRO CELL COUNTReduce analysis time and improve data accuracy

QUANTIFYOBSERVE

Image 1

AUTO MEASUREMENT &DATA ACCUMULATION

Image 2 to ××

ANALYSIS

Image 1

QUANTIFYOBSERVE QUANTIFYOBSERVE

Image 2

QUANTIFYOBSERVE

Image 3

QUANTIFYOBSERVE

Image ××

DATA SUMMARIZATION QUANTIFY

Since each image is measured separately, conditions may differ.

REDUCED TIME

MORE RELIABLE MEASUREMENTS IN LESS TIMEOUTPUT CONDITIONS• Threshold value• Correction values• Mask setting• Measurement target range (upper/lower limits)• Colocalization setting, etc

All images are measured under the same conditions, eliminating user bias.

DRAG & DROP

CONDITIONFILE

CONVENTIONAL METHOD

MACRO CELL COUNT

SAVE CONDITION

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KA12-1017The information in this publication is based on KEYENCE’s internal research/evaluation at the time of release and is subject to change without notice. Company and product names mentioned in this catalog are either trademarks or registered trademarks of their respective companies.The specifications are expressed in metric units. The English units have been converted from the original metric units.

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all-in-one fluorescence microscope – no darkroom …

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