Category Archives: NIKON A1R/TIRF/STORM

New automated diascopic shutter for DIC imaging on the A1R-setup

We bought a new shutter to rapidly block halogen light in time recordings using the EM-CCD camera of the NIKON imaging station. Previously, intervals between fluorescent observation and DIC recording required warming up the halogen lamp for at least 4 seconds. Now the light path is rapidly switchable in milliseconds and is incorporated into the NIS software and its Optical Configurations.

JOBS training

We had a training session at NIKON to work with the wizard creator ‘JOBS’ within the NIS software. It is aimed to easily guide people through an imaging session. Especially multiwell plate definition, random position imaging within wells, autofocus, different image capture for different wells  can be predefined and structured. Of course, users still need to know how to tweak the system to get good images but it may turn out to become a popular tool for our facility.

New Hard Drive to temporarily backup data, new Cy5 cube

The NIKON system has been updated again, a 2 TB hard drive is installed that allows temporary storage untill you verified dataintegrity . We will imply an automated sync-system  to move data from the C:\ drive to the new F:\ drive when the system is not in use.

A new cy5-cube has been placed in the 3rd A1R filter wheel set to allow sensitive imaging of far red dyes in the GaAsP detector, and outperforms the old filtercube several fold.

 

About imaging chambers

Users need to purchase their own stocks of imaging chambers, we were lucky to obtain a set of chambers from IBIDI and they perform well on our systems. We also have good experience with WPI 3 cm Fluodishes (having a 8 mm or 23 mm imaging well), initial problems with Labtech chambered coverglas slides are solved. Low cost solutions using ATTOfluor cell chambers clamping 25 mm round cs from Marienfeld are also available. It should be noted that mounting cs sometimes induces tension in the clamped cs which is released in an unpredictable manner, resulting in a loss of focus. Remember that objectives of high end imaging systems are calibrated to work with #1.5 (or 0.17 mm) thick glass coverslips.

Lastly, the TOKAI hit heater setup will be updated to allow better fluid replacement during imaging in the coming weeks.

Posts Before November 2013

dd October 2013

The A1R/Storm system is running  satisfactory, we keep testing its capacities. NIS software has been updated to the latest 4.2 version with quite some improvements. Also the SPE2 has been serviced and software was updated. Please contact us if you have difficulties.

dd June 2013

The NIKON A1R/STORM system is running. A1R CLSM live cell imaging is performing quite well, we await filter changes to optimally image gfp / rfp variants. The STORM is still in its testing phase, especially dual/triple color imaging is challenging. The AFM has moved to the CCI and is operational, contact Bart Gadella for further info.

dd April 2013

The NIKON A1R/STORM system has been installed and the first imaging validations are being preformed. Please ask staff if you are interested to image on the machine. We will evaluate the time schedule and need for you to do your experiment on the machine.

dd March 2013: 

A new state of the art microscope will be installed, the NIKON A1R/STORM system. We will be trained in the coming weeks on the A1R CLSM, the TIRF and STORM options.

The AFM from the B&C department will be moved to the CCI.

dd Jan 2013:

new Leica cryostate 3050S replaced the broken IRAS cryostate, thanks to collaborive effort of IRAS, DWM,Pathobiologie, I&I and GZD. Thank you all on behalf of the users for the continuation of this functionality at the CCI.

dd 2012

The OME image database on a LINUX server (with kind help of Aart deGroot) is working fine. We are exploring ways to properly implement it into the network to have central collection of the microscope data. We aim to store these for about half a year so users can acces the files easily and arrange definite storage capacity of their own.