Images produced by the light microscopy team at The Crick.

Light Microscopy : Latest news

News and updates about Crick Advanced Light Microscopy STP

Latest CALM updates:

New Confocal Arrivals: Installation of our new LSM 880 multi-photon system will begin the week of June 24th. The 880 MP will replace the 780 MP, which was moved to the 5th floor and remains open for general use as a CALM system. The new 880 MP is also inverted and will have a Ti:Saphire / OPO laser combination that expands imaging of multiple fluorophores through dual-line excitation and wavelength mixing. The following week we will take delivery on a new LSM 880 with AiryScan (also inverted). This workhorse will help take pressure of our existing 880 AiryScan system. Following the release of the new LSM 900 series in April (after we ordered our systems in December...) we are in discussion with Zeiss to upgrade the 880 Airyscan to a 980 in the near future. See Matt or Rocco for details.

Open Staff Positions: After 37 years working for the Crick, CRUK, and the ICRF, Trevor Duhig has moved on from his role as Senior Microscopist. We are currently advertising at the SLRS level for someone to fill Trevor’s position, which will involve a focus on confocal quality control. We also have approval now for a position at the LRS level to help with the management of confocal microscopes outwith the STP. This person will perform quality control checks, help manage repairs, and work closely with Trevor’s replacement to develop a QC strategy for confocal microscopes across the institute. See Kurt for details.

Data management: We now have a test version of OMERO running to evaluate this software for managing image data. Individual labs can use OMERO to facilitate the organisation, analysis, and sharing of image data. It has useful features for both researchers and lab heads. Our project to evaluate OMERO also has work packages dealing with Histopathology, High Throughput Screening, Long-term Time-lapse imaging, and analysis of STORM data using the high-performance cluster. See Kurt or Donald for details.

Huygens for AiryScan: SVI have released a Huygens module for deconvolution of AiryScan data which really improves on Zeiss’ own software. The Huygens software now accounts for the special geometry of the AiryScan detector. Anyone using the AiryScan for super-resolution should strongly consider processing their data using Huygens instead (for which we have a server license). See Dave or Matt for details.

Super-Resolution Demos: We currently have the OMX SR SIM system on extended demo. This system is suitable for imaging live and fixed tissue-culture cells and offers some advantages (as well as some disadvantages...) over the AiryScan. In July we will demo super-resolving spinning disk systems from Nikon and Olympus. See Matt or Donald for details.

Intravital Microscopy: Our new Olympus FV3000 multi-photon system has been installed in LB2-2681. This system offers several improvements over our LSM 700 MP system for intravital microscopy, including better correction for deep tissue imaging, extended laser tuning, and more sensitive detectors. The system is equipped with Olympus’ motorised 25x / 1.05 NA dipping objective which improves signal and resolution in tissue. It also has an Insight X3 laser with extended tuning range from 680 - 1300 nm for excitation of red fluorophores. Finally, the system is equipped with 4 non-descanned PMTs optimised for detection of scattered fluorescence, 2 of which are high-sensitivity GaAsP detectors. We are currently assembling the peripheral kit needed for intravital microscopy (temperature control, anaesthetic unit, dark box) and hope to start training users on this system soon. See Rocco or Donald for details.

Super-Resolution Microscopy: Many people have begun generating useful data with our Vutara STORM microscope. The system is proving easy to use for super-res newbies and has also received positive reviews from more experiences users. Todd Fallesen, our new image analyst, will provide specialised assistance for STORM data analysis. See Deb or Todd for details.

TIRF Microscopy: Our Olympus TIRF system is (finally!) operational. The system is equipped with 4 laser lines which can be individually adjusted for optimal evanescent field illumination (405, 488, 561, and 637 nm). See Donald or Kurt for details.

Training: We have just completed another lecture series on light microscopy, featuring weekly talks from CALM staff on topics including Intro to LM, Optical Sectioning, Confocal, Multi-photon, Light Sheet, and Live Cell Imaging. This series of talks is offered twice per year, and will be offered again starting in October. See Kurt or David Hudson for details.

 

Dr. Kurt I. Anderson
CALM Head
June 2019

Previous CALM updates:

Image Analysis: We have hired a second image analyst to work along side Dave Barry. Todd Fallesen is an alumni of the Surrey lab, and will start Monday, April 1st. Dave and Todd will be sitting in SW 302 along with Fabrice Prin (see below). They are available to work on any image analysis problems, not just light microscopy. See Dave for more info.

Senior Microscopist: We have hired a new senior microscopist to fill the post left open following Peter Jordan's retirement last year. Camille Charoy is currently a post-doc at UCL Ophthalmology and will start with us mid-April. She will start out supporting confocal microscopy, which will help free up Rocco to work on advanced methods including FLIM and FCS. See Rocco for more info.
 
Quadrant Confocal Quality Control: We will soon advertise for a person to perform quality control on confocal microscopes based in the quadrants. We currently perform weekly QC tests only on the 8 confocals in the facility, but our goal is to add bi-weekly testing for all confocals requested. See Kurt for more info.
 
Crick Imaging Workshop: We are planning an informal workshop on Monday, June 3rd in Auditorium 1 (10 - 4ish). The goal will be to share information about current capabilities, and assess future needs. The format will be short talks (10 min), ideally based around a specific question and how it might be addressed using imaging. We will include a few talks on advanced methods currently in use at the Crick, but the emphasis will be on discussing what people would like to do in the future (crazy thoughts welcome). This will help to identify areas of common interest and guide our investment in new methods and imaging systems. Attendance will be open to everyone. See Kurt or Donald for more info.
 
Data Management: We have started a project with Histopathology, High-Throughput Screening, and Scientific Computing to explore use of OMERO to manage image data across the Crick. OMERO makes it easier for people to organise, analyse, and share their data. Watch for training sessions beginning in the second half of April. See Kurt for more info.
 
HREM: High-resolution episcopic microscopy (HREM) is a serial block-face technique which can generate large ( >1 cm^3), high-resolution 3D volumes in the absence of specific fluorescent or histochemical signals. Its particularly useful for phenotyping mouse development. Fabrice Prin has developed this method working in Tim Mohun’s lab, and has now joined the CALM STP to continue supporting this service to the Crick. See Kurt or Fabrice for more info.
 
Intravital Microscopy: We are in the process of installing our new Olympus FV3000 multi-photon in B2-2681 for intra-vital microscopy. This system has a SpectraPhysics Insight X3 laser, which is tunable from 680 to 1300 nm, and an upright research stand for working with anaesthetised mice. The system has a number of advanced features, including a motorised collar which incrementally corrects for spherical aberration during deep Z-stacks, and an awesome 25x / 1.03 NA IR optimised objective. See Kurt, Donald, or Rocco for more details.
 
New confocals arriving: We have a number of new instruments arriving over the next couple of months. An LSM 880 with AiryScan has just been installed in a class-2 space in 4 NE. This instrument will be used heavily by the Tooze lab, but is also available for others needing open access to class-2 samples (you can image class-2 samples on some CALM systems in sealed chambers). We’re also expecting imminent delivery of a new LSM 880 with AiryScan in the STP. This system will replace one of our less-heavily used upright systems and has been specified primarily for super-resolution use. Finally, we’ll begin installation of our new LSM 880 MP system in early May. This system will have two MP lasers: a Chameleon Ultra II (tunable from 680 - 1080 nm @ 3 W peak power) and MPX OPO (1010 - 1340 nm @ 1 W peak power) and will be optimised for multi-spectral imaging. See Donald, Rocco, or Matt for details.
 
Super-resolution Demos: Our big purchase for 2019 will be a super-resolution system for live cell imaging, which will complement the STORM system (super-resolution for fixed samples) we bought last year. We will be looking at both SIM and super-res spinning-disk systems, and running demos through the spring into the summer. See Donald or Matt for details.
 
User Survey and Website: Thanks to everyone who responded to the recent STP user survey. We had 117 responses for light microscopy, the second highest response rate, and 60 comments. The comments were generally quite positive, although one theme which emerged both for us and across the STPs in general was that people felt they were not well informed about what’s going on in the STP (methods, technologies, events, etc). This is a fundamental challenge in an institute the size of the Crick and something we will aim to address in as many different ways possible (including this newsletter…). Our first goal will be to complete the re-vamp of our website which Rocco started a couple months ago, putting as much info as we can up there while keeping it as clear as possible. See Rocco for more info. 
 
Dr. Kurt I. Anderson
CALM Head
March 2019

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