AFM-FLIM facility

We recently installed and commissioned an integrated atomic force (AFM) and fluorescence lifetime imaging (FLIM) microscope, funded by an Strategic Equipment grant from EPSRC. More information coming soon.

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-UF3For 531 nm and/or 637 nm lasers
ZT 405-473/532 rpc-UF3For 402 nm and/or 531 nm lasers
ZT 488/532 rpc-UF3For 482 nm and/or  531 nm lasers
ZT 470-488/640 rpc-UF3For 482 nm and/or 637 nm lasers
ZT 405/485 rpc-UF3For 402 nm and/or 482 nm lasers
ZT 405 rdc-UF3For 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 CubeFor splitting all light into two directions (Polarisation independent)
H488lpxrReflected: λ < 488 nm Transmitted: λ > 488 nm
T635lpxrReflected: 475 nm < λ < 635 nm Transmitted: 650 nm < λ
BS R830Reflected: 350 nm < λ < 830 nm Transmitted: 845 nm < λ < 1600 nm (For Infrared detector)
Zt532rdc-UF1Reflected: 405 nm < λ < 532 nm Transmitted: 545 nm < λ < 750 nm

Filters

Blank (x17)For 100 % transmission
750 SPTransmitted: λ < 750 nm
425 LPTransmitted: λ > 425 nm
488 LPTransmitted: λ > 488 nm
532 LPTransmitted: λ > 532 nm
937 LPTransmitted: λ > 937 nm
445/30 BPTransmitted: 430 nm < λ < 460 nm
480/20 BPTransmitted: 570 nm < λ < 490 nm
511/20 BPTransmitted: 501 nm < λ < 521 nm
582/64 BPTransmitted: 550 nm < λ < 614 nm
690/70 BPTransmitted: 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).