This BYOC was designed to achieve one of my

This BYOC was designed to achieve one of my goals I had set upon joining the ISAC SRL EL program. The purpose of the BYOC is 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 the build list, instructions, notes and highlight there are many opportunities for improvement/modification (I can attest to this with my experience over the last 6 years). 

...

Notes - other references

mmHg to PSI

**Divide by 51.715 to obtain PSI**

...

38 pounds

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

$30 each 

or $30 for 2$38.50USD

Item	Qty	Purpose	Sources	Cost ($AUD)	Notes	Image
600x600 tile	1	base	Local tile supplier or Bunnings (requires a packet)	$0-$40 Free (tell them what you are making)	Aim for a free marble tile	Image Added
600x600 steel sheet	1	build platform	Bunnings (Sydney)	$26	Ensure it is magnetic. Thickness does not matter, thinner normally means cheaper. I recommend sign steel. Have some fun and wobble it ;p
0.3M7TIA gear	2	holds HPLC in the centre of luer lock	Aliexpress	<$5 for 20	https://www.aliexpress.com/item/33000449925.html?spm=a2g0s.9042311.0.0.86a84c4dqSRqUr
450 extension springs	19 (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 screw	bag	used as screws to put the whole thing together	RS components or ebay
M3 nut	bag	Ensure outside dimensions are 5.5mmx5.5mm	RS components	$13.80	RS PRO, M3 5.5mm Steel Square Nuts, Bright Zinc Plated Finish  837-262	Image Removed	25mmx25mm optics	2-3	focus light onto detector from intercept point	Edmund optic	$52.80	Stock #32-490 837-262	Image Added
25mmx25mm optics	2-3	focus light onto detector from intercept point	Edmund optic	$52.80	Stock #32-490 https://www.edmundoptics.com.au/p/25mm-dia-x-25mm-fl-mgfsub2sub-coated-double-convex-lens/2375/
5mmx25mm optics	1	focus laser light onto stream	Edmund optic	$61.50	Stock No. #87-913 https://www.edmundoptics.com.au/p/25mm50mm-dia-x-25mm250mm-fl-mgfsub2sub-coated-doubleplano-convex-lens/2375/	5mmx25mm optics	1	focus laser light onto stream	Edmund 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 nozzle	1	eject stream	Intertronics or FISNAR	/30185/
100um nozzle	1	eject stream	Intertronics or FISNAR	38 pounds	https://www.intertronics.co.uk/product/preci-tip-precision-dispensing-tips/ IDMSPREC-M100-NS
Luer lock connectors and 1/4inch BSP to barb fittings	1	to connect to nozzle	Bunnings & Kinesis Australia & USA KENT		I will need to separate this column a bit more - information will be provided during the workshop.	Image Added
30mm x 10mmx 3mm	20	mount items	Ebay	$18	Image Removed	30mm x 10mmx 3mm	20	mount items	Ebay	$18	20pcs N50 Strong Block Cuboid Rare Earth Neodymium Magnets 30mm x 10mm x 3mm https://www.intertronicsebay.cocom.ukau/product/preci-tip-precision-dispensing-tips/ IDMSPREC-M100-NS	Luer lock connectors and 1/4inch BSP to barb fittings	1	to connect to nozzle	Bunnings & Kinesis Australia & USA KENT	20pcs N50 Strong Block Cuboid Rare Earth Neodymium Magnets 30mm x 10mm x 3mmitm/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				Image Added
20L Drum	2	used to create a vacuum	recycle bin	free	to make see instructions
Regulator	2	adjust pressures	supercheap auto	$30 each  or $30 for 2	20L Drum	2	used to create a vacuum	recycle bin	free	to make see instructions	Regulator	2	adjust pressures	supercheap auto	https://www.supercheapauto.com.au/p/blackridge-air-regulator/340078.html or  https://www.ebay.com.au/itm/20pcs1-N504-StrongInch180PSI-BlockAir-CuboidCompressor-RarePressure-EarthRegulator-NeodymiumGauge-MagnetsSwitch-30mmControl-x-10mm-x-3mm/171206332711?hash=item27dcb15127:m:mkKfgJMNa9H2_6E0RsAkhxQ&frcectupt=true	Circular magnet	To mount APD instead of photodiode	Image Removed	Valve-Kits/264442958026?ssPageName=STRK%3AMEBIDX%3AIT&_trksid=p2057872.m2749.l2649
APD or Photodiode	1 minimum	convert photons to current	many sources including old instruments	$15 each	APDs used are AD500-8 TO52 package or BNC photodiode package
Laser 532nm (or equivalent)	1		AIXIZ lasers	$38.50USD	https://www.supercheapautoaixiz.com.au/p/blackridge-air-regulator/340078.html or  https://www.ebaystore/product-page/532nm-5-25mw-adjustable-power-3vdc-line-cross-attachable-ad-532-25adj
Electronics shopping list (per detector)
Prototyping board	1			$8.19	Laser 532nm (or equivalent)	1	AIXIZ lasers	https://core-electronics.com.au/itm/1solder-4-Inch180PSI-Air-Compressor-Pressure-Regulator-Gauge-Switch-Control-Valve-Kits/264442958026?ssPageName=STRK%3AMEBIDX%3AIT&_trksid=p2057872.m2749.l2649
APD or Photodiode	1 minumum	convert photons to current	many sources including old instruments	$15 each	APDs used are AD500-8 TO52 package or BNC photodiode package
able-breadboard.html
RJ45 breakout	2			$1.33	https://core-electronics.com.au/breakout-board-for-rj45.html	Image Added
LT1364CN8#PBF-ND	2	opamp used for I>V and for amplification	DIGIKEY	#13.73 each	https://www. aixiz digikey. store com/product- page detail/en/ 532nm linear- 5 technology- 25mw analog- adjustable-power-3vdc-line-cross-attachable-ad-532-25adj	Electronics shopping list (per detector)	Prototyping board	1	$8.19	https://core-electronics.com.au/solder-able-breadboard.html	RJ45 breakout	2	$1.33	https://core-electronics.com.au/breakout-board-for-rj45.html	Image Removed	LT1364CN8#PBF-ND	2	opamp used for I>V and for amplification	DIGIKEY	#13.73 each	https://www.digikey.com/product-detail/en/linear-technology-analog-devices/LT1364CN8-PBF/LT1364CN8-PBF-ND/891887	Cat7 RJ45 cable	1	$8	Amazon/Ebay	Adjustable 10^5 25 turn resistor devices/LT1364CN8-PBF/LT1364CN8-PBF-ND/891887 Now obsolete, any high speed high slew op amp with low noise, rail to rail should work. I will look to include a specific part soon.
Cat7 RJ45 cable	1			$8	Amazon/Ebay
Adjustable 10^5 25 turn resistor				<$1	Jaycar (generic electronics parts store)
Adjustable 10^3 25 turn resistor				<$1	Jaycar (generic electronics parts store)
1N4148 diodes	4			<$1	Jaycar (generic electronics parts store)
10pF ceramic capacitors	2			<$1	Jaycar (generic electronics parts store)
10K resistor (1% tolerance)	3			<$1	Jaycar (generic electronics parts store)
100K resistor	2			<$1	Jaycar (generic electronics parts store)
Adjustable 10^3 25 turn resistor1K resistor	1			<$1	Jaycar (generic electronics parts store)
1N4148 diodes22K resistor	41			<$1	Jaycar (generic electronics parts store)
10pF 0.22uF ceramic capacitorscap	21			<$1	Jaycar (generic electronics parts store)
10K resistor (1% tolerance)	31uF ceramic cap	1			<$1	Jaycar (generic electronics parts store)	100K resistor	2	<$1	Jaycar (generic electronics parts store)	1K resistor	1	<$1	Jaycar (generic electronics parts store)	22K resistor	1	<$1	Jaycar (generic electronics parts store)	0.22uF ceramic cap	1	<$1	Jaycar (generic electronics parts store)	1uF ceramic cap	1	<$1	Jaycar (generic electronics parts store)
2 pin molex pcb header 2.54mm pitch	1	$0.202 pin molex pcb header 2.54mm pitch	1			$0.20	https://www.jaycar.com.au/2-pin-0-1-straight-locking-header-2-54-pitch-single/p/HM3412
3pin molex header female 2.54mm pitch	1	used to connect to APD		$0.55	https://www.jaycar.com.au/3-pin-0-1-header-with-crimp-pins-2-54-pitch/p/HM3403	Image Added
hookup wire and crimpers				$0	Spares in every household
9V battery connectors	4	buy different types of connectors to ensure flat 80-90V power supply		$1.40	Jaycar
8 pin IC housing	2	mount for  IC chips		$1.25	https://www.jaycar.com.au/8-pin-gold-insert-machined-pin-ic-socket/p/PI6452
4 or 6 way DIP switch	1	used to turn power on / off		$1.65	https://www.jaycaraltronics.com.au/p/2s3050-pin4-0way-1-straight-locking-header-2-54-pitch-single/p/HM3412	3pin molex header female 2.54mm pitch	1	used to connect to APD	$0.55dip-switch/
shielded RJ45 port	2	needed to suppress noise		$1.75	https://www.jaycaraltronics.com.auau/p/3p1538-pinoupiin-08p8c-1rj45-headermodular-withsocket-crimpshielded-pins-2-54-pitch/p/HM3403	Image Removed	hookup wire and crimpers	$0	Spares in every household	9V battery connectors	4	buy different types of connectors to ensure flat 80-90V power supply	$1.40	8 pin IC housing	2	mount for  IC chips	$1.25pcb-mount/
Differential pressure regulation
pressure transducer - 24PCAFA6D *only required for digital pressure readout	1	accurate pressure differential between sample/flow cell	Element14	$61.89	https:// www au. jaycar element14.com .au/8-pin-gold-insert-machined-pin-ic-socket/p/PI6452	4 or 6 way DIP switch	1	used to turn power on / off	$1.65	https://www.altronics.com.au/p/s3050-4-way-dip-switch/	shielded RJ45 port	2	needed to suppress noise	$1.75	https://www.altronics.com.au/p/p1538-oupiin-8p8c-rj45-modular-socket-shielded-pcb-mount/	Differential pressure regulation
pressure transducer - 24PCAFA6D	1	accurate pressure differential between sample/flow cell	Element14	$61.89	https://au.element14.com/honeywell/24pcafa6d/pressure-sensor-0-1psid/dp/731638?st=24pc%20pressure%20sensor 0-1psi - 0-45mV - 2002508.pdf	Image Removed
OPAMP - breakout for pressure transducer	1	simple 2 stage amplification of 100	Core electronics	$9.90	BOB-09816  100x gain  https://core-electronics.com.au/opamp-breakout.html	Image Removed
LCD keypad	1	pressure output monitor	Core electronics	$16.50	https://core-electronics.com.au/lcd-keypad-shield-for-arduino.html DFR0009	Image Removed
arduino and case	1	arduino uno	Core electronics	$39.95 + $4.70	CE05634 A000066
Aluminum laser mount and heat sink	1	holds and cools the laser	Aixiz laser supplies	USD$3.95	12X30mm aluminum mount for AixiZ 12X30mm units [AIX-ALM-MT-1]	Image Removed
6mm nuts	many	mounts with 3mm threads	RS components	$12.23 pkt of 100	RS PRO, M3 5.5mm Steel Square Nuts, Bright Zinc Plated Finish 837-262	Image Removed
Molex 2 way connector and crimp	1x for laser power source	Allowing the disconnection renders the laser and power supply modular.	RS components	Image Removed	Cable ties	many	Need I say more	100mm x 2.5mm	Image Removed
Polulu fine control stepup/down  voltage regluator	1 per APD detector	9V batteries are coarse adjustment - this gets you finer control	core electronics	$27.15	https://www.pololu.com/product/2869 The output voltage of the regulator is controlled with a 12-turn precision potentiometer. Turning the potentiometer clockwise increases the output voltage, and it can be measured using a multimeter.	Image Removed

Instructions 

Setting up the laser

Most standard cytometers use a laser as a light source. The BYO cytometer is no different. Read on to mount your laser!

• Insert the laser of choice into the aluminum heat sink and hand tighten the screw to lock the laser in.
  Image Removed
• Here are 2 compatible lasers (left is a 405nm, right is a 532nm laser available from aixiz.com.
  Image Removed
• Insert 2x5.5mm square nuts into the laser mount nut sockets and attach the laser mount using 2xM3 10mm socket screws
  Image Removed
• Insert 2 rectangular magnets into each base to secure the laser to the steel base.
  Image Removed
• Attach a 2.5mm ~100mm cable tie to secure the laser power cables. This is to prevent them falling off with too much movement. 
  Image Removed
• Cut off the excess on all cable ties going forward
  Image Removed
• Crimp on a 2 way molex connector using a crimper
  Image Removed
• Attach the connector and use a scapel to expose the laser control potentiometer (the yellow heat shrink is what you need to cut back). 
  Image Removed
• The laser is now ready!

Making a flow cell...

• obtain male to male luer lock (transparent is great), and 2 small 0.3M7TIA gears
  Image Removed
• Setup the luer lock to hold the gears - this will be used to hold the PEEK tubing. 
  Image Removed
• Insert the precut peek tubing 
  Image Removed
• Connect the remaining luer lock connectors followed by a luer lock to barb connector as shown
  Image Removed
• Using 2 Y connectors, tubing and luer lock to bar connectors create the following Y connector that will join the stiff main flow cell. 
  Image Removed
• Pass the peek tubing up through the female luer lock connector.
• Create a sheath bleed point and transition to silicon tubing as shown.

Let's vacuum!

The instrument uses a vacuum to draw away excess liquid. There are a few variants to the vacuum (including a gravity version that I absolutely love, and not only because I designed it, and it lasts ~10 minutes).

This version is the electric vacuum pump version. 

...

/honeywell/24pcafa6d/pressure-sensor-0-1psid/dp/731638?st=24pc%20pressure%20sensor 0-1psi - 0-45mV - 2002508.pdf	Image Added
OPAMP - breakout for pressure transducer *only required for digital pressure readout	1	simple 2 stage amplification of 100	Core electronics	$9.90	BOB-09816  100x gain  https://core-electronics.com.au/opamp-breakout.html	Image Added
LCD keypad *only required for digital pressure readout	1	pressure output monitor	Core electronics	$16.50	https://core-electronics.com.au/lcd-keypad-shield-for-arduino.html DFR0009	Image Added
arduino and case *only required for digital pressure readout	1	arduino uno	Core electronics	$39.95 + $4.70	CE05634 A000066
Aluminum laser mount and heat sink	1	holds and cools the laser	Aixiz laser supplies	USD$3.95	12X30mm aluminum mount for AixiZ 12X30mm units [AIX-ALM-MT-1]	Image Added
6mm nuts	many	mounts with 3mm threads	RS components	$12.23 pkt of 100	RS PRO, M3 5.5mm Steel Square Nuts, Bright Zinc Plated Finish 837-262	Image Added
Molex 2 way connector and crimp	1x for laser power source	Allowing the disconnection renders the laser and power supply modular.	RS components			Image Added
Cable ties	many	Need I say more			100mm x 2.5mm	Image Added
Polulu fine control stepup/down  voltage regluator	1 per APD detector	9V batteries are coarse adjustment - this gets you finer control	core electronics	$27.15	https://www.pololu.com/product/2869 The output voltage of the regulator is controlled with a 12-turn precision potentiometer. Turning the potentiometer clockwise increases the output voltage, and it can be measured using a multimeter.	Image Added
2xD battery holder	2	Used to power the laser and vacuum pump	Jaycar	$2.75	https://www.jaycar.com.au/2-x-d-cell-side-by-side-battery-holder/p/PH9220 CAT.NO:  PH9220
Vacuum pump	1	Used to generate a vacuum	Core Electronics	$10.15	https://core-electronics.com.au/370-mini-vacuum-pump.html SKU: FIT0801 Brand: DFRobot Runs for over 24 hours using 2xD batteries. Current measured during use @ 3V = ~250mA.	Image Added

Instructions 

Getting started - making a base!

• Obtain a 600mm x 600mm tile (marble preferred but ceramic is more economical) 
• Place it on the left hand side of a table that is longer than the tile (we need the space on the right for the electronics).
  Image Added
• Place a 600mm x 600mm steel sheet over the tile. 
  Image Added
• Secure with clips as shown. 
  Image Added 

Air, sheath and vacuum reservoirs

• Place magnets, nuts and screws into the 3D printed bottle holders. 
  Image Added
  Image Added
• Place the 3 x bottle holders at the rear of the build as shown.
  Image Added

Let's vacuum!

The instrument uses a vacuum to draw away excess liquid. There are a few variants to the vacuum (including a gravity version that I absolutely love, and not only because I designed it, but because it uses gravity lasts ~10 minutes).

In 2022 we'll be setting up an battery operated electric vacuum pump. 

• Obtain a vacuum pump (listed above). 
  Image Added
• Solder the terminals to a 2xD size battery holder.
  Image Added
• Obtain a wine bottle with a aluminum lid, preferably a top shelf one . Utilise wine thoughtfully. 
• To drill the holes we start with a guide drill bit ~1-2mm, drill our holes while the lid is on the bottle and then follow through with a hole size that is about ~2mm smaller than the tubing. I've found a 4mm drill bit works well for the ~6mm Tygon tubing obtained for this project. The wine bottle lid requires 2 holes, 1 to draw out the vacuum and 1 to lead the liquid waste in.
  Image Added 
• Drill guide holes followed by larger holes (4mm for 6mm tubing)
  Image Added
  Image Added
• Insert tubing ends that have been cut on an angle to assist pulling tubing through. Lubrication with water of the tubing outer face may help. Ensure the tubing to be connected to the vacuum pump is cut close (within 1 inch to the bottle cap) to prevent liquid being sucked in to the hydrophobic filter. 
  Image Added
• Connect connectors including a hydrophobic filter to prevent liquid entering the vacuum pump. 

Setting up the laser

Most standard cytometers use a laser as a light source. The BYO cytometer is no different. Read on to mount your laser!

• Insert the laser of choice into the aluminum heat sink and hand tighten the screw to lock the laser in.
  Image Added
• Here are 2 compatible lasers (left is a 405nm, right is a 532nm laser available from aixiz.com.
  Image Added
• Insert 2x5.5mm square nuts into the laser mount nut sockets and attach the laser mount using 2xM3 10mm socket screws
  Image Added
• Insert 2 rectangular magnets into each base to secure the laser to the steel base.
  Image Added
• Attach a 2.5mm ~100mm cable tie to secure the laser power cables. This is to prevent them falling off with too much movement. 
  Image Added
• Cut off the excess on all cable ties going forward
  Image Added
• Crimp on a 2 way molex connector using a crimper
  Image Added
• Attach the connector and use a scapel to expose the laser control potentiometer (the yellow heat shrink is what you need to cut back). 
  Image Added
• The laser is now ready!

Making a flow cell...

• obtain male to male luer lock (transparent is great), and 2 small 0.3M7TIA gears
  Image Added
• Setup the luer lock to hold the gears - this will be used to hold the PEEK tubing. 
  Image Added
• Insert the precut peek tubing 
  Image Added
• Connect the remaining luer lock connectors followed by a luer lock to barb connector as shown
  Image Added
• Using 2 Y connectors, tubing and luer lock to bar connectors create the following Y connector that will join the stiff main flow cell. 
  Image Added
• Pass the peek tubing up through the female luer lock connector.
• Create a sheath bleed point and transition to silicon tubing as shown.