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Fluorescence Slide Scanning Microscope BUM400AFL


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  • Automated biological upright microscope
  • Motorized XY stage, Z focus, nosepiece
  • LED Fluorescence imaging for Dapi/Hoechst, GFP/FITC/Alexa 488, TRITC/mCherry, plus optional custom filter such as red or Infrared.
  • Sharp autofocus at custom size of steps, number of steps and skipped or focused spots.
  • Plan 4x, 10x, 20x objective lenses, optional 1.25x, 2x, 2.5x, 5x, 25x, 40x, 50x.
  • Scanning of 50x75mm double-size glass slide or two single slide 25x75mm
  • Preview of tiled /stitched image
  • Direct access to individual images, save at standard formats.
  • No limit on the number of tile size

$19,995.00 $45,000.00

BUM400AFL Automated Fluorescence Slide Scanning Microscope

BUM400AFL is our recent product designed specifically for scanning of a single or double glass slide in brightfield and epi-fluorescence imaging with optional darkfield and polarized imaging. Reflected mode is also available upon request.

Why BUM400AFL? This vs competitor products in the market.

There are two popular whole slide-scanning microscopes in the market. Both are made by two brands of Zeiss and Leica. Their price is at 6 digits. Ours are way below that and you will save at least 50% in the price. We understand our microscope is not as fast as those products, but its quality of images remains reasonably great. On top of that we offer this at small packages, which is affordable for small businesses and new PIs in the universities. Our winning point is that lots of customization can be offered such as custom filter sets per your application, camera resolution and sensitivity per your selection and range of objective lenses.

Features

  • Automated biological upright epi-fluorescence microscope
  • Motorized XY stage, Z focus, nosepiece
  • LED Fluorescence imaging for Dapi/Hoechst, GFP/FITC/Alexa 488, TRITC/mCherry, plus optional custom filter such as red or Infrared.
  • Sharp autofocus at custom size of steps, number of steps and number of skipped spots.
  • Plan 4x, 10x, 20x objective lenses, optional 1.25x, 2x, 2.5x, 5x, 25x, 40x, 50x, 100x.
  • Scanning of 50x75mm double-size glass slide or two single slide 25x75mm
  • Preview of tiled /stitched image
  • Direct access to individual images, save at standard formats (TIFF, JPEG, BMP, PNG, GIF, …).
  • No limit on the number of tile size
  • Optional advanced image analysis for measurement and tracking
  • Optional automatic slide loader (racks of 100, 200,…).

BUM400AFL Automated Fluorescence Slide Scanning Microscope

The BUM400AFL is an automated upright biological microscope with LED fluorescence imaging specifically designed for scanning of fluorescent glass slides or any fluorescent samples can be visualized from top view.

BUM400AFL is our recent product designed specifically for scanning of a single or double glass slide in brightfield and epi-fluorescence imaging with optional darkfield and polarized imaging. Reflected mode is also available upon request.

The system includes by default 4x, 10x, and 20x objective lenses but it can be upgraded easily or selected from the beginning with other lenses. Low magnification lenses are great for quick scan of the whole slide view. For this purpose, we have 1.25x, 2x, 2.5x, 4x and 5x objective lenses. It happens the customer struggles deciding between 20x and 40x objective lenses, as 20x has less resolution but 40x requires at least four times more scanning time of 20x lens. For this type of application, we suggest using 25x objective lens, which is at middle point between 10x and 40x. If you need a high magnification over 40x, you can select 50x (dry or oil), 60x (dry or oil), or 100x (oil).

Whole Slide Imaging

For whole slide imaging, a 2x obj lens does a great job.

 

Field of Views

This table shows the field of view size (Length x height) in micrometers x micrometers using a 2.0MP (1920×1080) Camera and a 0.65x C-Mount adapter:

Objective Lens
Field of View
Pixel Size (um)
2.0x5,000 um x 2,813 um2.604
2.5x4,400 um x 2,475 um2.292
4x2,438 um x 1,371 um1.270
10x988 um x 556 um0.515
20x510 um x 287 um0.266
40x233 um x 131 um0.121

Speed

Time for image acquisition and image processing (stitching the tiles together and displaying) depends on the sample size  and imaging condition. The number of tiles or mosaics depends on the sample size and what objective lens is selected, as well as the individual field of view (FOV) size. An FOV size itself is determined by the camera sensor sensor size and the c-mount adapter magnification.

Imaging condition includes the type of imaging (brightfield, fluorescnce, darkfield, etc), light intensity, exposure time (and gain determined by the camera type) and most importantly using autofocus condition. Auto-focusing is not required at low magnifications such as 2x, 2.5x, 4x or 5x. If you use a thin sample, this may not be still required at 10x obj lens. For 10x, you may choose partial autofocusing, for instance every 3, 5 or 10 spots of view. Depending on the sample type and if a high magnification such as 20x or above is selected, you may need to do the autofocuisng at each FOV. Meanwhile, the focus range and the number of steps for autofocusing , as well as speed of focusing is matter.

Prior running a big experiment, it is highly recommended to spend enough time to find the right setting parameters for acquisition such as exposure time and focusing parameters. Run small size of experiments, watch the result, tune the parameters then increase the whole size of experiments.

Here is a list of examples to get an idea of the speed using BUM400AFL.

Objective LensSpot SizeTile SizeImaging Condition
Spent Time(Acquisition Time + processing Time = Total (min: sec)
Speed (Area/time,mm^2 / second)Time (seconds /number of tiles)
1.25x27mm  x 72mm (whole slide size)9×6brightfield,

AE, Non AF

01:35 +0:10=01:4518.51.94
2x25.3mm x75mm (whole slide size)15×9

=135

brightfield

autoexposure

03:14 +00:25

=03:39

8.561.56
2.5x25mm x75mm (whole slide size)16 x 11

=176

brightfield

AE

06:37 + 00:36

= 07:13

4.332.46
2.5x20mm x22mm

(Coverslip)

5 x 8 =40brightfield

AE

01:26 +00:08=

01:34 (94 sec)

4.682.35
4xa large sample

(Spinach)

12mm x 8mm

5 x 6 =30Green fluorescence

ME

00:52+ 00:04

=00:56

1.711.87
10x12mm x 8mm11 x 14 = 154fluorescence, ME, AF (1 FOV, 10×10 = 100 steps)11:12+ 00:25

= 11:37

0.1454.53

Notes: AE: autoexposure, ME: Manual Exposure, AF: Autofocus

High Power LED Fluorescence Attachment

The microscope use a 3W LED for UV, blue and green channels. Custom LEDs are available.

Technical Specification

1. Microscope Optics

 DESCRIPTIONSPECIFICATIONIncluded
Optical SystemBasic UnitBUM400A Automated Microscope1
Control SystemBUM400A-CS for XY stage and Z Focus1
Eyepiece tube30°hinged, binocular, (55mm-75mm)1
ObjectivesPlan 4x, 10x, 20x, 40x1
Revolving nosepieceCan hold up to 4 objectives1
EyepiecesWF 10×/ 222
CondenserN.A 1.25 Abbe condenser with variable diaphragm and filter1
Power Adapter110V or 220 V1
FocusFine/Coarse coaxial focus to use gear rack transmission mechanism with 0.002 mm resolution1
Filter1
LampHigh brightness Kohler illumination, Adjustable Halogen lamp 12V/20W (or its equal brightness LED)

AC 85V – 230V

1
Serial/USB data cableBUM400A Automated Microscope exclusive1
StageTravel  range: 50x75mm

Speed: 1 to 50 mm/sec

1
ComputerIntel Core i5 or higher / 500 GB HDD/DVD / 8GB

with 19’’ LED

Optional
Others
Slide Loaderautomatic slide loader for racks of 100, 200 slidesOptional

 

2. Fluorescence Module

LED*FluorophoresFilters
NameEXNameEXEMEXDMEM
Ultraviolet (UV)365nmDapi / Hoechst350 nm470 nm330-385nm400420-470nm
Blue light (B)470nmFITC / GFP490 nm525 nm460nm-495nm505510-550
Green light (G)505nmAlexa Fluor 555555 nm580 nm460nm-550nm570590-650
Red (R)

(Optional)

625nmAlexafluor 647650 nm665 nm600-660nm660660nm
Custom400-1000nmvisible, IR

* The specs of LED and filters may change per the customer request.

** Ask us to customize the LEDs and Filters among  our over 40 filter sets.

We also have 5W LED source. Please check this product BIA-FL-IS5.

3. Camera

a) CMOS Cameras

Order CodeSensor &

Size (mm)

Pixel (μm)G Sensitivity
Dark Signal
FPS/ResolutionBinningExposure
BIC-E3S-2.02.0MP

IMX385(C)

1/2″(7.20×4.05)

3.75 x3.752350mv with 1/30s
0.15mv with 1/30s
125 @1920×10801×10.1ms ~15s
BIC-E3C-2.32.3MP

IMX185(C)

1/1.9″

(7.20×4.50)

3.75×3.751120mv with 1/30s
0.5mv with 1/30s
[email protected]×1200
[email protected]×600
1×1,

2×2

0.244ms~2s
BIC-E3C-2.3G2.3MP

IMX249 (C,GS )
1/1.2“

(11.25×7.03)

5.86×5.861016mv with 1/30s
0.15mv with 1/30s
[email protected]×12001×10.244ms~2s
BIC-E3S-3.1G3.1MP

IMX265 (C, GS)

1/1.8″(7.07×5.30)

3.45×3.451146mv with 1/30s
0.15mv with 1/30s
53 @2048×1536

85 @1024×768

1×10.1ms ~15s
BIC-E3S-5G5.0MP

IMX264(C, GS)
2/3” (8.45×7.07)

3.45×3.451146mv with 1/30s
0.15mv with 1/30s
35 @2448×2048
50 @1224×1024
1×10.1ms ~15s
BIC-E3S-9.0G9M

IMX305 (C, GS)

1″(14.13×7.45)

3.45 x3.451146mv with 1/30s
0.15mv with 1/30s
[email protected]×2160
[email protected]×1080
1×1,
2×2,
0.1ms ~15s
BIC-E3S-20.020MP

IMX183(C)

1″

(13.06×8.76)

2.4 x 2.4462mv with 1/30s
0.21mv with 1/30s
[email protected]×3648
50 @2736×1824
[email protected]×1216
1×1,
2×2,
3×3
0.1ms~15s

Note: C: Color, M: Monochrome, GS: Global Shutter

b) CCD, please visit this link: https://www.bioimager.com/product-category/cameras/ccd-cameras/2-stage-cooling/

c) sCMOS, Please visit this link: https://www.bioimager.com/product-category/cameras/scmos/

 

Software

1. User friendly interface

The slide scanning software has a very easy to use and user friendly  graphical user interface (GUI):

 

2. Easy and Simple Automation

The software has a very simple interface. You can move the XY stage, Z focus, autofocus at three speeds of movement (low, medium and high) with a click of mouse. It automatically rotates the obj lens, shows the light intensity, coordinates of the current position (X,Y,Z) and saves 3 custom positions for scanning, start or stop.

3. Move and position your sample

A sample can be moved and positioned at three different speeds of low, medium and high which can be set at custom values in all directions of horizontally (X), vertically (Y) and in height or depth of focus (Z).

4. Automatic Scanning

Depending on which objective lens is selected, you can define, select and save  the speed of movement, focusing range (number of focus, step size), step size (in X and Y direction), number of tiles (in X and Y), number  of autofocusing, a pause time at each field of view.

You can select fast scan without autofocusing.

You can also determine to reset the focus when you rotate the nosepiece and move between objective lenses or shut down the system.

5. Custom Objective Lens name, Light Intensity and Coordinates

The software allows easily to rename the objective lenses, change their position, define a default light intensity for that specific objective lens. In below example, we often switch between 2.5x and 40x. The system remembers that is the same spot but it should apply a different light intensity.

For safety reasons, you do have access to change the limit of movements in all directions of X, Y and Z. You can also choose to release or not the focus height during the rotate of  the nosepiece.

Videos & Images:

A) Videos

A1. This video demonstrates an example of  fluorescence (in red channel) imaging of a potato thin-section slide using 20x objective lens. It overview the software setting for this case study, shows autofocusing, manual exposure and also reviews the final image after stitching 18×21 tiles.

 

B) Images

Whole Coverslip 20mm x 20mm Imaging

A 20x22mm area is scanned by 2.5x obj lens with a 5×8 tile and processed in less than 94 seconds (1min 34 sec).

 

Example 1. Brightfield Imaging of a cartilage tissue slide

1.a. Using a 4x obj lens, and 5×3 tile:

1.b. Same Slide with 10x obj lens and 9×5 tiles

1.c. Cartilage Slide scanned with 20x obj lens, 18×9 tiles, manual exposure, autofocus every 5 spots

 

Example 2. Potato thin Section Slide

2.1. Brightfield imaging using 4x objective lens, with 4×5 tiles, autoexposure, non autofocus.

2.2. Fluorescence imaging using 4x objective lens, with 4×5 tiles, manual exposure 600ms,  autofocus on every 3 spots with focus range of 10 steps of 40=400 steps (each step has 0.31um height).

2.3. Brightfield imaging using 10x objective lens, with 9×11 tiles, autoexposure,  autofocus on every 3 spots with focus range of 16 steps of 8= total 128 steps (each step has 0.31um height).

2.4. Fluorescence imaging using 10x objective lens, with 9×11 tiles, autoexposure,  autofocus on every 1 spots with focus range of 16 steps of 8= total 128 steps (each step has 0.31um height).

2.5. Fluorescence imaging using 20x objective lens, with 19×20 tiles, manual exposure,  non autofocus.

 

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