A step-by-step guide to building a prism-type TIRF microscope
Total Internal Reflection Florescence (TIRF) Microscopy is a technique used to observe a very thin region of sample directly at the surface of an interface (coverslip) without obtaining background interference from the rest of the sample volume.
Dalton Gibbs, Anisa Kaur, Anoja Megalathan, Kumar Sapkota, and Soma Dhakal have published a wonderful paper entitled "Build Your Own Microscope: Step-By-Step Guide for Building a Prism-Based TIRF Microscope".
As the title implies, it is a step-by-step guide to build a prism-based TIRF microscope. It includes a bill of materials, step-by-step instructions, software links, beautiful diagrams and a level of technical detail that goes far beyond just how to build the device. We love to see this kind of thing.
The bill of materials doesn't have any prices, so we thought we'd put together as many specific prices as we could here, which come to a total of around $30,000. This includes everything and we do mean everything, from the table to the optical glasses. You can save money in a lot of ways by swapping out parts like the shutters.
| Catalog # | Description | Quantity | Vendor | Cost |
| p/n 325–1206 | p/n 325–1206 UV FS Pellin-Broca Prism 11 × 20 × 6.4 mm | 2 | Altos Photonics, Inc. (Bozeman, MT, USA ) | $242 |
| zet640/20x | magnetron bandpass clean-up filter (excitation path), 0–5 deg aoi | 1 | Chroma Technology Corp (Bellows Falls, VT, USA) | $325 |
| t565spxxr-uf3 | magnetron shortpass dichroic, 45 deg aoi | 1 | Chroma Technology Corp | $475 |
| 1069417 | SYS: CUBE 640-40 CIRCULAR: 640 nm: 40 mW | 1 | Coherent Inc. (Santa Clara, CA, USA) | $450 |
| 1073840 | ASSY: HEAT SINK: ACCESSORY: CUBE | 1 | Coherent Inc. | $400 |
| 1214333 | Productivity Plus Bronze-CUBE | 3 | Coherent Inc. | |
| EW-06419-01 | Tygon Microbore Autoanalysis Tubing, 0.020” × 0.060”OD, 100 ft/roll | 4 | Cole-Parmer | $228 |
| CL532-050-L | 532 nm Central Wavelength 50 mW CW Power | 1 | CyrstaLaser (Reno, NV, USA) | |
| 4001 | Hardman DOUBLE/BUBBLE Extra-Fast Set Epoxy Red Package 3.5 g Packet | 3 | Ellsworth Adhesive Systems (Germantown, WI, USA) | $300 |
| KBH-5503 | Laser Protective Eyewear for HeNe Alignment and KTP Alignment applications | 2 | Kentek Corporation (Pittsfield, NH, USA) | $400 |
| 9470 | 15 ft USB 3.0 A Male to A Female Active Extension Cable | 2 | Monoprice (Rancho Cucamonga, CA, USA) | $33 |
| 5010 | Cat6 24AWG UTP Ethernet Network Patch Cable, 20 ft Gray | 2 | Monoprice | $9 |
| 2067 | USB to RS232 DB9 male(Serial)/DB25 male Converter Cable | 1 | Monoprice | $10 |
| U-R380 | IX3-D6RES;6-POSITION IX NOSEPIECE CODED, DIC | 1 | Olympus America Inc. (Center Valley, PA, USA) | |
| U-V111C | U-TV1XC;C-MOUNT CAMERA ADAPTER, CENTERABLE | 1 | Olympus America Inc. | |
| 9-U734 | 45FR; 45MM FROSTED DIFFUSION FILTER, IX3 | 1 | Olympus America Inc. | |
| UYCP-11 | UYCP-11;US STYLE 3-PRONG POWER CORD | 1 | Olympus America Inc. | |
| 5-UR403 | IX3-RFA;STRAIGHT ILLUMINATOR | 1 | Olympus America Inc. | |
| 5-UR416-1 | IX3-RFACS-1-2; CODED IX3FLUORESCENCE TURRET | 1 | Olympus America Inc. | |
| OCT-TD7BX3 | TRF59907-OL3; Dual-band ET-532/640 nm laser TIRF set | 1 | Olympus America Inc. | |
| OAT-DU-897U-CS0-#BV | DU-897U-CS0-#BV; IXON ULTRA897 EMCCD, 56FPS, 512 × 512, 16 UM, USB | 1 | Olympus America Inc. | |
| OAT-TR-EMFS-F06 | 532/640 EM SPLITTING 3 PART FILTER SET FOR DPC&OPTOSPLIT | 1 | Olympus America Inc. | |
| OAT-TR-DCIS-CA1-00 | DUAL CAM CASSETTE. REQ(TR-EMF S-F) | 1 | Olympus America Inc. | |
| O89-OptoIILS | OptoSplit II LS–1.0x; Optosplit II system w/cubes and diaphragm | 1 | Olympus America Inc. | |
| OMT-010 | MT-010; LOGO CLOTH DUSTCOVER 11 × 25 × 26”, ANTI-STATIC BX, IX | 1 | Olympus America Inc. | $35 |
| M-ST-UT2-46-8; Tuned-Damped Table, 1200 × 1800 × 203 mm, M6 Holes | 1 | $10,000 | ||
| OVP-S2000A428 | S-2000A-428; 28” Isolators w/Auto Leveling, Set of 4 | 1 | Olympus America Inc. | |
| OVP-ACWS | ACWS; Air Compressor, Low Noise, 110 V | 1 | Olympus America Inc. | $704 |
| Computer & Monitor | $800 | |||
| DIB-551.00 | Diamond Coated “Stick” Drills, DIB-551.00 | 10 | Shor International | $90 |
| TR6-P5 | Ø1/2” Optical Post, SS, 8-32 Setscrew, 1/4”−20 Tap, L = 6”, 5 Pack | 3 | Thorlabs, Inc. (Newton, NJ USA) | $33 |
| MB1218 | Aluminum Breadboard 12” × 18” × 1/2”, 1/4”−20 Taps | 1 | Thorlabs, Inc. | $194 |
| SH8S050 | 8-32 Stainless Steel Cap Screw, 1/2” Long | 1 | Thorlabs, Inc. | $7 |
| TR12 | Ø1/2” Optical Post, SS, 8-32 Setscrew, 1/4”−20 Tap, L = 12” | 6 | Thorlabs, Inc. | $68 |
| SH25S038 | 1/4”-20 Stainless Steel Cap Screw, 3/8” Long | 1 | Thorlabs, Inc. | $8 |
| RA90 | Right-Angle Clamp for Ø1/2” Posts, 3/16” Hex | 8 | Thorlabs, Inc. | $80 |
| TR6 | Ø1/2” Optical Post, SS, 8-32 Setscrew, 1/4”−20 Tap, L = 6” | 1 | Thorlabs, Inc. | $7 |
| TR075 | Ø1/2” Optical Post, SS, 8-32 Setscrew, 1/4”−20 Tap, L = 0.75” | 2 | Thorlabs, Inc. | $10 |
| SS25S075 | 1/4”−20 Stainless Steel Setscrew, 3/4” Long, Pack of 25 | 1 | Thorlabs, Inc. | $7 |
| SH25S075 | 1/4”−20 Stainless Steel Cap Screw, 3/4” Long, Pack of 25 | 1 | Thorlabs, Inc. | $9 |
| B3648F | 36” × 48” × 2.4” Imperial Breadboard, 128 × 98 × 23 cm | 1 | Thorlabs, Inc. | $1,876 |
| PSY313 | 900 × 1200 mm Full Under Shelf, 146 × 95 × 6 cm | 1 | Thorlabs, Inc. | $259 |
| PTA512 | Air Compressor-110/115 V-60 Hz, US Power Plug, 45 × 38 × 46 cm | 1 | Thorlabs, Inc. | $662 |
| RSP1 | Rotation Stage For 1” Optics 2.2”OD 1.062-20 ID | 1 | Thorlabs, Inc. | $91 |
| TR3-P5 | 1/2” Dia. × 3” Length: Pack of 5 Post | 3 | Thorlabs, Inc. | $75 |
| SS6MS25 | M6-1.0 × 25 mm Set Screw, 25 Pack | 2 | Thorlabs, Inc. | $17 |
| SS6MS12 | M6 × 1.0 Stainless Steel Set Screw 12 mm Long Pack of (25) | 1 | Thorlabs, Inc. | $6 |
| FMP1-P5 | Fixed Ø1” Optical Mount 5-Pack | 2 | Thorlabs, Inc. | $158 |
| WP25M-VIS | Mounted Ø25.0 mm Wire Grid Polarizer, 420–700 nM | 1 | Thorlabs, Inc. | $291 |
| ID8 | Mounted Standard Iris, 8.0.mm max. Aper. | 1 | Thorlabs, Inc. | $51 |
| ID12 | Iris Diaphragm 1/2” | 1 | Thorlabs, Inc. | $51 |
| ID15 | Mounted Standard Iris, 15.0.mm max. Aper. | 1 | Thorlabs, Inc. | $51 |
| WPMH05M-532 | Mounted Multi Order 1/2 Waveplate 532 nm | 1 | Thorlabs, Inc. | $260 |
| BB1-E02 | Ø25.4 mm Mirror, Broadband 400–750 nm | 4 | Thorlabs, Inc. | $310 |
| LB1904-A-ML | Mounted N-BK7 Bi-Convex Lens, Ø1”, f = 125 mm, -A | 1 | Thorlabs, Inc. | $49 |
| LB1437-A-ML | Mounted N-BK7 Bi-Convex Lens, Ø1”, f = 150 mm, -A | 1 | Thorlabs, Inc. | $48 |
| LB1945-A-ML | Mounted N-BK7 Bi-Convex Lens, Ø1”, f = 200 mm, -A | 1 | Thorlabs, Inc. | $48 |
| ESK01 | MOUNTING SUPPORTS ESSENTIALS KIT #1 | 1 | Thorlabs, Inc. | $818 |
| LG1 | Laser Glasses, 190–400 nm, 808–1090 nm | 1 | Thorlabs, Inc. | $189 |
| ADB-10 | Pellin Broca Prism 10 mm BK7 | 1 | Thorlabs, Inc. | $75 |
| MT3A/M | XYZ Metric Translator Stage | 1 | Thorlabs, Inc. | $1,240 |
| KS1 | Lockable Kinematic 1” Optic Mount | 4 | Thorlabs, Inc. | $93 |
| SDA90120S | Standing Height Active Science Desk to suit 900 × 1200 mm | 1 | Thorlabs, Inc. | $2,822 |
| LS6S2T0 | Uni-stabled housed Shutter | 1 | Vincent Associates | $795 |
| VCM-D1 | Shutter Driver | 1 | Vincent Associates | $1,210 |