SPIM

A Compact and Versatile Light Sheet Microscopy Platform for Research and Education


Selective Plane Illumination Microscope (SPIM) with sample rotation & translation. Optimized for uncompromised light sheet microscopy of samples like Drosophila embryos and spheroids while maintaining a small footprint for mobility and a reasonable price point. Epi-fluorescence and brightfield illumination, 16x/0.8 W objective, sCMOS camera. Most of the parts are off-the-shelf, with only three custom components made by Selective Laser Sintering (SLS).

Light sheet properties

 * Scheme of the light sheet illumination arm with laser (blue), lenses (gray), mirrors (brown) and detection (green). Incoming laser beam is expanded to 2.7 mm diameter.


 * 491 nm laser beam in water as seen in the field of view (FOV) after passing through the illumination arm. Cylindrical lens, emission filter and dichroic mirror removed. 22 ms exposure time. Full width at half maximum (FWHM) determined using Gaussian fitting in Fiji. Yellow lines indicate ideal Gaussian shape.

Parts

 * C = custom part / part that requires modification
 * some categories require parts from the Basics and Lenses category
 * tools not included
 * US prices from 2016/2017

Illumination arm
10X Nikon CFI Plan Fluorite Objective, 0.30 NA, 3.5 mm WD	Nikon		1	 $-  	 $-   	illumination objectives ($1,315.60 @ Thorlabs)	https://www.thorlabs.com/thorproduct.cfm?partnumber=N10XW-PF Cobolt Calypso 491nm 100mW DPSS	Cobolt		1	 $-  	 $-   	491 nm laser (from old laser launch)	http://www.cobolt.se/product/04-01-series/ NEOS PCAOM	NEOS	48062-2.5-.55	1	 $-  	 $-   	AOTF (from old laser launch) NEOS 4-channel PCAOM driver	NEOS	64040-75-.1.4CH	1	 $-  	 $-   	AOTF controller (from old laser launch) Adapter with External SM1 Threads and Internal M25 x 0.75 Threads	Thorlabs	SM1A12	1	 $19.75 	 $19.75 	adapter illumination objective to SM1	http://www.thorlabs.com/thorproduct.cfm?partnumber=SM1A12 25.4mm Diameter, 4-6λ Mirror	Edmund Optics	#69-245	4	 $16.00 	 $64.00 	1" mirror, only 3 mm thick, for gimbal mount	http://www.edmundoptics.com/optics/optical-mirrors/flat-mirrors/4-6-wave-first-surface-mirrors/69245 Z-Axis Translation Mount, 30 mm Cage Compatible	Thorlabs	SM1Z	1	 $186.00 	 $186.00 	zoom housing for telescope lens	https://www.thorlabs.com/thorproduct.cfm?partnumber=SM1Z 30mm Cage System Adjustable Slit, 8-32 Tap, Imperial Micrometer	Thorlabs	VA100C	2	 $260.00 	 $520.00 	slit for setting light sheet properties, 30 mm cage system	http://www.thorlabs.com/thorproduct.cfm?partnumber=VA100C Kinematic Mount with Vertical Drive, Ø1/2" Optics	Thorlabs	VM05	4	 $97.90 	 $391.60 	vertical drive 0.5" mirror mounts for laser module	https://www.thorlabs.com/thorproduct.cfm?partnumber=VM05 Ø1/2" Broadband Dielectric Mirror, 400 - 750 nm	Thorlabs	BB05-E02	4	 $49.40 	 $197.60 	0.5" mirrors for laser module	https://www.thorlabs.com/thorproduct.cfm?partnumber=BB05-E02 30 mm Cage System Iris, Ø20.0 mm Maximum Aperture	Thorlabs	CP20S	8	 $85.00 	 $680.00 	iris for beam alignment, 30 mm cage system	http://www.thorlabs.com/thorproduct.cfm?partnumber=CP20S Ø1/2" Optical Post, SS, 8-32 Setscrew, 1/4"-20 Tap, L = 1.5"	Thorlabs	TR1.5	8	 $4.97 	 $39.76 	shorter posts for laser mounts	https://www.thorlabs.com/thorproduct.cfm?partnumber=TR1.5 Kinematic Mount with Vertical Drive, Ø1/2" Optics	Thorlabs	VM05	4	 $97.90 	 $391.60 	0.5" mirror mounts w/ vertical drive	https://www.thorlabs.com/thorproduct.cfm?partnumber=VM05 Unmounted Continuously Variable ND Filter, Ø50 mm, OD: 0-4.0, ARC: 350 - 700 nm	Thorlabs	NDC-50C-4-A	1	 $368.00 	 $368.00 	ND filter wheel for laser module	https://www.thorlabs.com/thorproduct.cfm?partnumber=NDC-50C-4-A Post Mount Assembly for Round, Unmounted, ND Filters	Thorlabs	NDC-PM	1	 $52.25 	 $52.25 	mount for ND filter wheel	https://www.thorlabs.com/thorproduct.cfm?partnumber=NDC-PM Assmann WSW Components H7VXH-0906M	Digi-Key	H7VXH-0906M-ND	2	 $5.18 	 $10.36 	cable for triggering AOTF	http://www.digikey.com/product-detail/en/assmann-wsw-components/H7VXH-0906M/H7VXH-0906M-ND/998782 Pomona Electronics 4969	Digi-Key	501-1030-ND	4	 $8.99 	 $35.96 	cable for BNC input to other cable	http://www.digikey.com/product-detail/en/pomona-electronics/4969/501-1030-ND/603340

Detection arm
16X Nikon CFI LWD Plan Fluorite Objective, 0.80 NA, 3.0 mm WD	Nikon		1	 $-  	 $-   	detection objective ($5,824.21 @ Thorlabs)	https://www.thorlabs.com/thorproduct.cfm?partnumber=N16XLWD-PF Infinity-Corrected Tube Lens for Plan Fluorite Objectives	Nikon		1	 $-  	 $-   	Nikon 200 mm tube lens for detection ($450 @ Thorlabs)	https://www.thorlabs.com/thorproduct.cfm?partnumber=ITL200 Camera	Hamamatsu	Orca Flash 2.8	1	 $-  	 $-   	main camera	http://www.hamamatsu.com/jp/en/community/life_science_camera/product/search/C11440-10C/index.html BP 525/50, mounted, 25 mm	Chroma	ET525/50m	1	 $-  	 $-   	filter for EX 491 nm, GFP etc. ($325 @ Chroma)	https://www.chroma.com/products/parts/et525-50m 8-Channel, 16-Bit Analog Voltage Output Device	MCC	USB-3103	1	 $349.00 	 $349.00 	USB-to-analog-out for AOM etc.	http://www.mccdaq.com/usb-data-acquisition/USB-3103.aspx Adapter with External SM1 Threads and Internal M32 x 0.75 Threads	Thorlabs	SM1A34	1	 $22.00 	 $22.00 	adapter detection objective to SM1	http://www.thorlabs.com/thorproduct.cfm?partnumber=SM1A34 Adapter with External SM2 Threads and Internal M38 x 0.5 Threads	Thorlabs	SM2A20	1	 $45.00 	 $45.00 	tube lens to SM2 adapter	https://www.thorlabs.com/thorproduct.cfm?partnumber=SM2A20 60 mm Cage Plate, SM2 Threaded, Enhanced Clamping, 0.5" Thick, 8-32 Tap	Thorlabs	LCP08	1	 $47.90 	 $47.90 	60 mm cage plate w/ SM2 thread	https://www.thorlabs.com/thorproduct.cfm?partnumber=LCP08 60 mm Cage Plate with Ø2" Double-Bore Optic Mount, 8-32 Tap	Thorlabs	LCP06	1	 $33.50 	 $33.50 	60 mm cage plate for holding SM2 tube	https://www.thorlabs.com/thorproduct.cfm?partnumber=LCP06 30 mm to 60 mm Cage Plate Adapter, 8-32 Tap	Thorlabs	LCP02	2	 $38.00 	 $76.00 	60-to-30 mm cage adapter	http://www.thorlabs.com/thorproduct.cfm?partnumber=LCP02 Adapter with External C-Mount Threads and External SM1 Threads	Thorlabs	SM1A39	1	 $20.00 	 $20.00 	C-mount-to-SM1 adapter	https://www.thorlabs.com/thorproduct.cfm?partnumber=SM1A39 Adapter with External SM2 Threads and Internal SM1 Threads	Thorlabs	SM2A6	1	 $24.00 	 $24.00 	SM1-to-SM2 adapter	https://www.thorlabs.com/thorproduct.cfm?partnumber=SM2A6 SM2 Lens Tube Without External Threads, 1/2" Thread Depth, Two Retaining Rings Included	Thorlabs	SM2M05	1	 $24.50 	 $24.50 	SM2 tube for cage plate	https://www.thorlabs.com/thorproduct.cfm?partnumber=SM2M05 60 mm Cage Plate with Ø2.2" Double Bore for SM2 Lens Tube Mounting	Thorlabs	LCP09	1	 $41.20 	 $41.20 	60 mm cage plate for SM2 tube	https://www.thorlabs.com/thorproduct.cfm?partnumber=LCP09 Dual-Position Slider Bundle: Dual-Position Slider, Interface Board, Cables	Thorlabs	ELL6K	1	 $268.00 	 $268.00 	motorized filter switch	https://www.thorlabs.com/thorproduct.cfm?partnumber=ELL6K SM1 Lens Tube, 0.30" Thread Depth, One Retaining Ring Included	Thorlabs	SM1L03	3	 $12.16 	 $36.48 	filter mount for motorized filter switch	https://www.thorlabs.com/thorproduct.cfm?partnumber=SM1L03 30 mm to 60 mm Cage Plate Adapter, 8-32 Tap	Thorlabs	LCP02	3	 $38.00 	 $114.00 	cage mount for filter switch, wheel	https://www.thorlabs.com/thorproduct.cfm?partnumber=LCP02 Cage Assembly Rod, 2" Long, Ø6 mm, 4 Pack	Thorlabs	ER2-P4	1	 $22.57 	 $22.57 	cage extension rods	https://www.thorlabs.com/thorproduct.cfm?partnumber=ER2-P4 Cage Assembly Rod, 1" Long, Ø6 mm, 4 Pack	Thorlabs	ER1-P4	2	 $18.77 	 $37.54 	cage rods for mounting switch and wheel	https://www.thorlabs.com/thorproduct.cfm?partnumber=ER1-P4 SM1 Lens Tube, 1.00" Thread Depth, SM1RR Retaining Ring, 5 Pack	Thorlabs	SM1L10-P5	1	 $67.70 	 $67.70 	SM1 tubes for detection path	https://www.thorlabs.com/thorproduct.cfm?partnumber=SM1L10-P5

Extra illumination
Cold White Mounted LED, 1000 mA, 800 mW (Min)	Thorlabs	MCWHL5	1	 $193.00 	 $193.00 	LED for brightfield illumination	https://www.thorlabs.com/thorproduct.cfm?partnumber=MCWHL5 Blue (470 nm) Mounted LED, 1000 mA, 650 mW (Min)	Thorlabs	M470L3	1	 $268.00 	 $268.00 	LED for widefield EX 470 nm	https://www.thorlabs.com/thorproduct.cfm?partnumber=M470L3 T-Cube LED Driver, 1200 mA Max Drive Current	Thorlabs	LEDD1B	2	 $293.00 	 $586.00 	LED driver	https://www.thorlabs.com/thorproduct.cfm?partnumber=LEDD1B 15 V, 2.4 A Power Supply Unit for One K-Cube or T-Cube	Thorlabs	KPS101	3	 $25.71 	 $77.13 	power supply for LED driver	https://www.thorlabs.com/thorproduct.cfm?partnumber=KPS101 BP 470/40, mounted, 25 mm	Chroma	ET470/40x	1	 $-  	 $-   	clean-up filter for EX w/ 470 nm LED ($325 @ Chroma)	https://www.chroma.com/products/parts/et470-40x DC 488, unmounted, 36x25 mm	Chroma	ZT488rdc	1	 $-  	 $-   	dichoric for EX w/ 470 nm LED ($300 @ Chroma)	https://www.chroma.com/products/parts/zt488rdc SM1 Lens Tube, 1.00" Thread Depth, One Retaining Ring Included	Thorlabs	SM1L10	2	 $14.25 	 $28.50 	SM1 tube for blocking LED stray light	https://www.thorlabs.com/thorproduct.cfm?partnumber=SM1L10 30 mm Cage Plate, Ø1.2" Double Bore for SM1 Lens Tube Mounting	Thorlabs	CP12	2	 $20.00 	 $40.00 	cage plate for mounting LED	https://www.thorlabs.com/thorproduct.cfm?partnumber=CP12