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Build Your Own Cytometer - ACS

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This BYOC was designed as one of my goals of the ISAC SRL EL program to act as an additional aid to spark interest when teaching cytometry to students. It has been designed to be affordable ($500-$1500), 3D printable and fun to build. I have included build list and instructions and highlight there are many opportunities for improvement/modification. 

The BYOC Workshop was first held @ ACS 2016 in Sydney, Australia, @ ACS 2018 in Adelaide, Australia and conjointly with the Garvan Flow Facility @ ACS 2019, Melbourne, Australia. 

Check back for updates. 

Overview

The cytometer uses a <$50 adjustable 5-25mW laser (interchangeable) that is directed onto a jet in air stream. The stream is ejected throughout a 100um nozzle that is assembled using luer locks, cogs and HPLC tubing to allow injection of sample in a laminar flow. The laser is focused onto the stream and signals are detected using a 25mm, 25mm focal lens that directs the light onto the FSC/SSC/FL detectors. A diagram of the light path from excitation is shown in the diagram below. The signals are detected with a APD/Photodiode or PMT and then converted to a voltage output using a pre-amp. The signals can be viewed using an oscilloscope or can be displayed on a bivariate dot plot using a ADC connected to a computer. 

3D Printable Parts

Steering laser mirror mount - https://www.thingiverse.com/thing:3523131

Flow cell - https://www.thingiverse.com/thing:3902321

Detector - https://www.thingiverse.com/thing:3902266

Mounting - https://www.thingiverse.com/thing:3902335

Sample tube mount - https://www.thingiverse.com/thing:3902328

Flow control - https://www.thingiverse.com/thing:3902432

Laser mount - https://www.thingiverse.com/thing:3902366

Bill of materials - 3D Printed Version (acrylic assists can be interchanged)

ItemQtyPurposeSourcesCost ($AUD)NotesImage
600x600 tile1baseLocal tile supplierFree (tell them what you are making)Aim for a free marble tile (smile)
600x600 steel sheet1build platform Bunnings (Sydney)$26Ensure it is magnetic. Thickness does not matter, thinner normally means cheaper. I recommend sign steel. Have some fun and wobble it ;p
0.3M7TIA gear2holds HPLC in the centre of luer lockAliexpress<$5 for 20

https://www.aliexpress.com/item/33000449925.html?spm=a2g0s.9042311.0.0.86a84c4dqSRqUr

450 extension springs19 (2 detectors), additional 6 per detector provides pulling force to mirror, laser, lenses

Bolts & Industrial (Sydney)

Century Springs (US)

$4

We need to buy it from a reseller.

Spring Catalogue: https://www.centurysprings.com.au/products/prod_catalogue.htm

Can also buy about 10x cheaper from ebay.


M3 socket screwbagused as screws to put the whole thing togetherRS components or ebay


M3 nutbagEnsure outside dimensions are 5.5mmx5.5mmRS components$13.80RS PRO, M3 5.5mm Steel Square Nuts, Bright Zinc Plated Finish 
837-262

25mmx25mm optics2-3focus light onto detector from intercept pointEdmund optic$52.80

Stock #32-490

https://www.edmundoptics.com.au/p/25mm-dia-x-25mm-fl-mgfsub2sub-coated-double-convex-lens/2375/


5mmx25mm optics1focus laser light onto streamEdmund optic$61.50

Stock No. #87-913

https://www.edmundoptics.com.au/p/50mm-dia-x-250mm-fl-mgfsub2sub-coated-plano-convex-lens/30185/


100um nozzle1eject streamIntertronics or FISNAR38 pounds

https://www.intertronics.co.uk/product/preci-tip-precision-dispensing-tips/

IDMSPREC-M100-NS


Luer lock connectors and 1/4inch BSP to barb fittings1to connect to nozzleBunnings & Kinesis Australia & USA KENT 

I will need to seperate this column a bit more - information will be provided during the workshop. 


30mm x 10mmx 3mm20mount itemsEbay $18

20pcs N50 Strong Block Cuboid Rare Earth Neodymium Magnets 30mm x 10mm x 3mm

https://www.ebay.com.au/itm/20pcs-N50-Strong-Block-Cuboid-Rare-Earth-Neodymium-Magnets-30mm-x-10mm-x-3mm/171206332711?hash=item27dcb15127:m:mkKfgJMNa9H2_6E0RsAkhxQ&frcectupt=true



Circular magnet 
To mount APD instead of photodiode


20L Drum2used to create a vacuum recycle binfreeto make see instructions
Regulator2adjust pressuressupercheap auto

$30 each 

or $30 for 2

https://www.supercheapauto.com.au/p/blackridge-air-regulator/340078.html

or 

https://www.ebay.com.au/itm/1-4-Inch180PSI-Air-Compressor-Pressure-Regulator-Gauge-Switch-Control-Valve-Kits/264442958026?ssPageName=STRK%3AMEBIDX%3AIT&_trksid=p2057872.m2749.l2649


APD or Photodiode 1 minumumconvert photons to currentmany sources including old instruments$15 each

APDs used are AD500-8

TO52 package or BNC PD package

  
Laser 532nm (or equivalent)1
AIXIZ lasers$38.50USDhttps://www.aixiz.store/product-page/532nm-5-25mw-adjustable-power-3vdc-line-cross-attachable-ad-532-25adj







Electronics shopping list (per detector)





Prototyping board1

$8.19

https://core-electronics.com.au/solder-able-breadboard.html


RJ45 breakout2

$1.33https://core-electronics.com.au/breakout-board-for-rj45.htmlBreakout Board for RJ45 BOB-00716 Sparkfun Australia - Express Delivery Australia Wide (Image 6)
LT1364CN8#PBF-ND2opamp used for I>V and for amplificationDIGIKEY#13.73 eachhttps://www.digikey.com/product-detail/en/linear-technology-analog-devices/LT1364CN8-PBF/LT1364CN8-PBF-ND/891887
Cat7 RJ45 cable1

$8Amazon/Ebay
Adjustable 10^5 25 turn resistor


<$1Jaycar (generic electronics parts store)
Adjustable 10^3 25 turn resistor


<$1Jaycar (generic electronics parts store)
1N4148 diodes4

<$1Jaycar (generic electronics parts store)
10pF ceramic capacitors2

<$1Jaycar (generic electronics parts store)
10K resistor (1% tolerance)3

<$1Jaycar (generic electronics parts store)
100K resistor2

<$1Jaycar (generic electronics parts store)
1K resistor1

<$1Jaycar (generic electronics parts store)
22K resistor1

<$1Jaycar (generic electronics parts store)
0.22uF ceramic cap1

<$1Jaycar (generic electronics parts store)
1uF ceramic cap1

<$1Jaycar (generic electronics parts store)
2 pin molex pcb header 2.54mm pitch1

$0.20https://www.jaycar.com.au/2-pin-0-1-straight-locking-header-2-54-pitch-single/p/HM3412
3pin molex header female 2.54mm pitch1used to connect to APD
$0.55https://www.jaycar.com.au/3-pin-0-1-header-with-crimp-pins-2-54-pitch/p/HM3403
hookup wire and crimpers


$0Spares in every household
9V battery connectors4buy different types of connectors to ensure flat 80-90V power supply
$1.40

8 pin IC housing2mount for  IC chips
$1.25https://www.jaycar.com.au/8-pin-gold-insert-machined-pin-ic-socket/p/PI6452
4 or 6 way DIP switch1used to turn power on / off
$1.65https://www.altronics.com.au/p/s3050-4-way-dip-switch/
shielded RJ45 port2needed to suppress noise
$1.75https://www.altronics.com.au/p/p1538-oupiin-8p8c-rj45-modular-socket-shielded-pcb-mount/










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