AFM-FLIM facility – Inaugural event

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

We are holding an inaugural event on Wednesday 14th April (on Microsoft Teams) and it is open for everyone interested. We will share details of the kit and ways to access it, and we have two great invited speakers who will tell us how AFM and FLIM systems can be harnessed to obtain further insights in materials science. Please register for the event here.

The schedule is as follows:

10.00-10.15 Introduction to equipment (Dr Nacho Martin-Fabiani)
10.15-11.15 Dr Ioanna Mela (University of Cambridge)
11.15-12.15 Dr Marina Kuimova (Imperial College London)
12.15-12.30 Access and final remarks (Dr Nacho Martin-Fabiani)

Dr Ioanna Mela (University of Cambridge)Transient behaviour of nanomechanical domains in triple cation perovskite films studied by multimodal microscopy

Dr Ioanna Mela gained her PhD in Pharmacology at the University of Cambridge and is currently a Research associate in the Laser Analytics Group, in the Department of Chemical Engineering and Biotechnology at the University of Cambridge. Ioanna has worked with high-speed and conventional atomic force microscopy (AFM) and she is currently developing and using correlative imaging platforms such as AFM/SIM, AFM/STED and AFM/FLIM. She is interested in applications of correlative AFM both in biology and in materials science. Beyond correlative AFM, her research interests are focused on developing DNA nanostructures that can specifically interact with bacterial targets while carrying active antimicrobials and she is bringing together different microscopy techniques to understand, characterise and quantify those interactions, with the view to develop antimicrobial drug delivery vehicles

Dr Marina Kuimova (Imperial College London)- Mapping microscopic viscosity and crowding using molecular rotors and FLIM

Marina Kuimova is a Reader (Associate Professor) at Imperial College London. Her current research is focused on elucidation of biologically relevant processes using different types of fluorescence imaging and time-resolved spectroscopy. She is a Fellow of the Royal Society of Chemistry and a member of the Editorial Board of Methods and Applications of Fluorescence. She has received numerous awards and honors for her work, including 2011 Grammaticakis-Neumann Prize of the Swiss Chemical Society, 2009 Roscoe the Westminster Medals at the SET for Britain, UK Houses of Parliament; 2012 British Biophysical Society Young Investigator Award, 2012 Royal Society of Chemistry Harrison-Meldola Prize, 2013 ChemComm Emerging Investigator Lectureship, the 2014 IUPAP C6 Young Scientist Prize in Biological Physics and • Society of Porphyrins and Phthalocyanines Young Investigator Award, 2020.

Marina obtained her Master’s Degree at Moscow State University (Russia), and a doctorate at the University of Nottingham (UK) under the supervision of Professor M. W. George in 2006. Following a postdoctoral appointment with Professor David Phillips at Imperial, she became a group leader and an EPSRC Life Science Interface Fellow (in 2007) and an EPSRC Career Acceleration Fellow (in 2010). She was appointed as a lecturer in the department of Chemistry at Imperial in 2012 and promoted to a Readership in 2016.

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-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).