We recently installed and commissioned an integrated atomic force (AFM) and fluorescence lifetime imaging (FLIM) microscope, funded by an Strategic Equipment grant from EPSRC.
SYSTEM SPECIFICATIONS
AFM system: Bruker BioScope Resolve
Tapping, contact, PeakForce QNM, and ScanAsyst modes.
Can measure topography, nanomechanics, and conductivity/IV curves (under development).
FLIM system: PicoQuant Microtime 200
Laser sources: LDH-D-C-405 (405 nm), LDH-D-C-485 (485 nm),LDH-D-TA-530B (532 nm), and LDH-D-C-640 (640 nm)
Objectives (Olympus):
Visible range: Plan N 20x NA 0.40 (air), Plan N 40x NA 0.65 (water), PlanFL N 100x / NA 0.95 (air) WD = 0.2 mm, UPlanSApo 60x NA 1.20 (water) WD = 0.25 mm
Infrared: LCPlan N 20x NA 0.45 IR WD = 8.3 mm; LCPlan N 100x NA 0.85 IR WD = 1.48 – 1.18 mm (glass)
Epifluorescence filters: DAPI, eGFP, mCherry TexaRed
Major dichroic mirrors
| ZT 532/640 rpc-UF3 | For 531 nm and/or 637 nm lasers |
| ZT 405-473/532 rpc-UF3 | For 402 nm and/or 531 nm lasers |
| ZT 488/532 rpc-UF3 | For 482 nm and/or 531 nm lasers |
| ZT 470-488/640 rpc-UF3 | For 482 nm and/or 637 nm lasers |
| ZT 405/485 rpc-UF3 | For 402 nm and/or 482 nm lasers |
| ZT 405 rdc-UF3 | For 402 nm laser |
Beam splitter optics
| 100% Mirror (x4) | For reflecting 100 % of the incoming light |
| 50/50 BS Plate (x4) | For splitting all light into two directions (Polarisation dependent) |
| 50/50 BS Cube | For splitting all light into two directions (Polarisation independent) |
| H488lpxr | Reflected: λ < 488 nm Transmitted: λ > 488 nm |
| T635lpxr | Reflected: 475 nm < λ < 635 nm Transmitted: 650 nm < λ |
| BS R830 | Reflected: 350 nm < λ < 830 nm Transmitted: 845 nm < λ < 1600 nm (For Infrared detector) |
| Zt532rdc-UF1 | Reflected: 405 nm < λ < 532 nm Transmitted: 545 nm < λ < 750 nm |
Filters
| Blank (x17) | For 100 % transmission |
| 750 SP | Transmitted: λ < 750 nm |
| 425 LP | Transmitted: λ > 425 nm |
| 488 LP | Transmitted: λ > 488 nm |
| 532 LP | Transmitted: λ > 532 nm |
| 937 LP | Transmitted: λ > 937 nm |
| 445/30 BP | Transmitted: 430 nm < λ < 460 nm |
| 480/20 BP | Transmitted: 570 nm < λ < 490 nm |
| 511/20 BP | Transmitted: 501 nm < λ < 521 nm |
| 582/64 BP | Transmitted: 550 nm < λ < 614 nm |
| 690/70 BP | Transmitted: 655 nm < λ < 725 nm |
Detectors
Single Photon Counting Module Excelitas SPAD (x2): Red sensitive 400- 1000 nm.
Photomultiplier module PMA Hybrid 40 (x2): Response 300 – 720 nm.
NIR-PMT detector: Spectral range 950 – 1700 nm.
TCSPC electronics: Hydraharp (high time resolution, for fast lifetimes) and MultiHarp (for longer lifetimes).
Martin-Fabiani Group 