Facility Head:
Dr Anand JEYASEKHARAN
CSI Singapore
Facility Head:
Dr Anand JEYASEKHARAN
CSI Singapore
The Light Microscopy Facility in CSI is equipped with a variety of microscopes to support the research requirements of the institute. The facility spans across the entire three levels of CSI and contains several imaging systems with different designs and configurations to meet variable research application requirements. Access to the microscopes is contingent on adherence to rules concerning appropriate usage of equipment. All new users must undergo training conducted by facility staff before they are allowed to access the facility.
Microscopy is fundamentally important to the field of molecular biology as it provides researchers with quantitative and spatial information on cellular structures at the micron to sub-micron level. The light microscope facility in CSI is geared towards providing researchers with the ability to acquire this information through a holistic approach which capitalises on the equipment readily available within the institute. The facility is available to staff and students of CSI for multi-mode high-resolution digital imaging and image analysis. It consists of a wide variety of microscopes ranging from confocal microscopes to wide field epi-fluorescence microscopes for different research applications. A separate workstation equipped with Metamorph, Nikon NIS Elements AR and ImageJ/Fiji is available for off-line image processing and analysis.
This is a sample workflow for general microscopy:
The Light Microscopy Facility currently houses the following equipment:
Nikon A1R Confocal – 13 South confocal room
The Nikon A1R is a modular confocal system consisting of a separate scan head, standard 4 PMT detector module plus an additional PMT for pseudo-DIC imaging, control module, laser module, and spectral detector module. The A1R has a standard high-resolution confocal mode (up to 4096 x 4096 pixels) as well as a high speed resonant mode capable of scanning 230 frames per sec at 512 x 64 pixels. In addition to laser-based, confocal image acquisition, this Nikon platform also utilizes a high-end digital camera for non-confocal image acquisition.
Features:
Filter sets:
Filter set 1 (DAPI, Hoechst, AlexaFluor 405, Pacific Blue, Cascade Blue, Coumarin) | Filter set 2 (FITC, GFP, eCFP) | Filter set 3 (TRITC, AlexaFluors 546, 551, 568, Cy3, DiI, DsRed, mCherry, PE, PI, RFP, tdTomato,) | Filter set 4 (Cy5, Alexafluors 633, 647, To-PRO-3) | |
Ex/Em | 330 – 380 nm / 420 nm | 465 – 495 nm / 515 – 555 nm | 540/25 nm / 605/55 nm | 590-650 nm / 663-738 nm |
Beamsplitter | 400 nm | 505 nm | 565 nm | 660 nm |
Objectives:
Laser Module (Class 3b):
Detector:
Software:
Applications:
Nikon C1 Confocal – 13 South confocal room
The Eclipse C1 Plus is a modular confocal imaging system that gives you the highest-quality digital images with an ultra-compact and lightweight design. The improved scan head optics of the C1 Plus boost optical performance to 400nm and its bi-directional scanner provides faster image acquisition. The system also includes support for X and Y scan axes rotation and new laser options for improved control over laser illumination intensity.
Features:
Filter Sets:
Filter set 1 (DAPI) | Filter set 2 (CFPHQ) | Filter set 3 (FITC) | Filter set 4 (Cy3) | Filter set 5 (YFPHQ) | |
Ex/Em | 340 – 380 nm / 435 – 485 nm | 420 – 445 nm / 460 – 510 nm | 465 – 495 nm / 515 – 555 nm | 512 – 552 nm / 565 – 615 nm | 490 – 500 nm / 520 – 560 nm |
Beamsplitter | 400 nm | 450 nm | 505 nm | 512 nm | 510 nm |
Objectives:
Laser Module (Class 3b):
Detector:
Software:
Applications:
Zeiss Axio Z1 with Cell Observer – 13 North Live Imaging room
Features:
Filter Sets:
Filter set 62 HE B/G/HR (DAPI, GFP, HcRed) | Filter set 43 (Cy3, DsRed) | Filter set 46 (YFP) | Filter set 47 ( CFP) |
|
Ex/Em | EX BP 370/40BP 474/28BP 585/35EM DBP 425/46+527/54+LP 615 | EX BP 545/25EM BP 605/70 | EX BP 500/20EM BP 535/30 | EX BP 436/20EM BP 480/40 |
Beamsplitter | BS TFT 395+495+610 | FT 570 | FT 515 | FT 455 |
Objectives:
Fluorescence Illumination:
Camera:
Software:
Applications:
Zeiss Axio Imager M2 – 11 North microscope room
Features:
Filter Sets:
Filter set 02 (DAPI) | Filter set 03 (FITC) | Filter set 04 (Cy3) | Filter set 05 (Cy5) | Filter set 06 (CFP) | |
Ex/Em | Ex BP 365Em LP 397 | Ex BP 450 – 490Em BP 515 – 565 | Ex BP 546Em LP 590 | Ex BP 640/30Em BP 690/50 | Ex BP 436/20Em BP 480/40 |
Beamsplitter | FT 395 | FT 510 | FT 580 | FT 660 | 455 |
Objectives:
Fluorescence Illumination: X-Cite Series 120 Q
Camera: Photometrics CoolSnap HQ2 monochrome CCD camera with 1392 x 1040 imaging array (6.45 x 6.45-?m pixels)
Software: Microsoft Windows 7 Pro with Zeiss ZenBlue (Dell Optiplex 3010 with Intel Core i5, Win 7 and 1TB HDD)
Applications:
Zeiss Axiovert A1 – 11 North microscope room
Features:
Filter Sets:
Filter set 02 (DAPI) | Filter set 03 (FITC) | Filter set 04 (Rhodamine) | Filter set 05 (EGFP) | |
Ex/Em | BP 365 / LP 397 | BP 450 – 490 / BP 515 – 565 | BP 546 / LP 590 | BP 470 / BP 525 |
Beamsplitter | FT 395 | FT 510 | FT 580 | FT 495 |
Objectives:
Fluorescence Illumination: X-Cite Series 120 Q
Camera: Photometrics CoolSnap HQ2 monochrome CCD camera with 1392 x 1040 imaging array (6.45 x 6.45-?m pixels)
Software: Microsoft Windows 7 Pro with ZenBlue (Dell Optiplex 3010 with Intel Core i5, Win 7 and 1TB HDD)
Applications:
Zeiss Axioplan 2 – 12 North microscope room
Features:
Filter Sets:
Filter set 02 (DAPI) | Filter set 03 (FITC) | Filter set 04 (Rhodamine) | Filter set 05 (EGFP) | |
Ex/Em | BP 365 / LP 397 | BP 450 – 490 / BP 515 – 565 | BP 546 / LP 590 | BP 470 / BP 525 |
Beamsplitter | FT 395 | FT 510 | FT 580 | FT 495 |
Objectives:
Fluorescence Illumination:
Camera: Zeiss AxioCam HrC (color image capture at 3900 x 3090 pixel resolution)
Software: Microsoft Windows XP with Zeiss AxioVision 4.6.3
Applications:
Olympus IX71 – 12 North microscope room
Features:
Filter Sets:
Filter set WU (DAPI) | Filter set WIBA (FITC, GFP) | Filter set WIGA ( TRITC) | |
Ex/Em | 360 – 370nm / 420nm | 460 – 490 nm / 515 nm | 515 – 550 nm / 600 – 640 nm |
Beamsplitter | 400 nm | 505 nm | 580 nm |
Objectives:
Fluorescence Illumination:
Camera:
Software: Microsoft Windows Vista with DP Controller
Applications:
Olympus SZX12 – 12 North microscope room
Features:
Filter Sets:
Filter set G | Filter set GFP | |
Ex/Em | 460 – 560nm / 590nm | 460 – 490 nm / 510 nm |
Objectives:
Fluorescence Illumination:
Camera:
Software:
Applications:
Zeiss PALM LCM – 13 North histology room
Features:
Objectives:
Laser Modules:
Detector:
Software:
Applications:
Nikon Eclipse TE2000S – 12 North microscope room
Features:
Filter Sets:
Filter set UV (DAPI) | Filter set B (FITC, GFP) | Filter set G (TRITC) | |
Ex/Em | 330 – 380 nm / 420 nm | 465 – 495 nm / 515 – 555 nm | 540/25 nm / 605/55 nm |
Beamsplitter | 400 nm | 505 nm | 565 nm |
Objectives:
Fluorescence Illumination:
Camera:
Software:
Applications:
Nikon TS100 – 13 North live imaging room
Features:
Filter Sets
Filter set B (FITC, GFP) | Filter set G (TRITC) | |
Ex/Em | 450 – 490 nm / 520 nm | 510 – 560 nm / 590 nm |
Beamsplitter | 505 nm | 575 nm |
Objectives:
Fluorescence Illumination:
Camera:
Software:
Applications:
190mm high stage (when the condenser is removed) allows for imaging of cell cultures at the bottom of a tall flask or testing chamber
Perkin Elmer Vectra Multispectral Imager – 13 North histology room
Features:
Objectives:
Fluorescence Illumination:
Camera:
Software:
Applications:
Zeiss Axiovert 200 – 12 North microscope room
Features:
Filter Sets:
Filter set 02 (DAPI) | Filter set 03 (FITC) | Filter set 46 (YFP) | Filter set 47 ( CFP) |
Filter set 04 (Rhodamine) | |
Ex/Em | BP 365 / LP 397 | BP 450 – 490 / BP 515 – 565 | Ex BP 500/20Em BP 535/30 | Ex BP 436/20Em BP 480/40 | BP 546 / LP 590 |
Beamsplitter | FT 395 | FT 510 | FT 515 | FT 455 | FT 580 |
Objectives:
Fluorescence Illumination:
Camera:
Software:
Applications:
Leica DM2500 Multiview with 5 stations – 12 North microscope room
Features:
Objectives:
Applications:
One PC workstation for offline image analysis
o Intel Core i7 5820K 3.3GHz 6-core processor
o 128GB SSD (Boot Drive) & 1TB HDD (Storage Drive)
o 32GB Memory
o AMD R9 290X Graphics Card for 3D visualisation
o ImageJ/Fiji
o Metamorph 7.7.7
o Nikon NIS Elements AR offline
o Zen Blue 2012 Lite
o Cell Profiler
Cancer Science Institute of Singapore
Centre for Translational Medicine
14 Medical Drive, #12-01,
Singapore 117599