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. It takes 3-4minutes to scan whole slide. If you use a 1.25x obj lens this will be even way better. It takes just 1-2 minutes to complete the full scan.
A fluorescence slide for mercury/HBO light alignment with green fibers inside.
Scanning area with BUM400AFL can be the full size of the glass slide or just the coverslip.
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). Below is the raw image with tiled boarder lines.
Please check the “imaging” tab for further sample images.
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:
Field of View
|Pixel Size (um)|
|2.0x||5,000 um x 2,813 um||2.604|
|2.5x||4,400 um x 2,475 um||2.292|
|4x||2,438 um x 1,371 um||1.270|
|10x||988 um x 556 um||0.515|
|20x||510 um x 287 um||0.266|
|40x||233 um x 131 um||0.121|
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, fluorescence, 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 Lens||Spot Size||Tile Size||Imaging Condition|
|Spent Time(Acquisition Time + processing Time = Total (min: sec)|
|Speed (Area/time,mm^2 / second)||Time (seconds /number of tiles)|
|1.25x||27mm x 72mm (whole slide size)||9×6 =54||brightfield,|
AE, Non AF
|2x||25.3mm x75mm (whole slide size)||15×9|
|2.5x||25mm x75mm (whole slide size)||16 x 11|
|06:37 + 00:36|
|5 x 8 =40||brightfield|
01:34 (94 sec)
|4x||a large sample|
12mm x 8mm
|5 x 6 =30||Fluorescence|
|10x||12mm x 8mm||11 x 14 = 154||fluorescence, ME, AF (1 FOV, 10×10 = 100 steps)||11:12+ 00:25|
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 and 5W power are also available.