Center for Microscopy

Putting Advanced Technology to Work

The Bigelow Analytical Services (BAS) Facility is located on Bigelow Laboratoryís Ocean Science and Education Campus in East Boothbay, Maine. The state≠-of-the-≠art facility provides analytical services to public and private entities. This advanced technology is available to local, national, and international researchers in all fields of study including marine chemistry, aquaculture, pharmacy, and fisheries. Our experienced scientists and technicians provide the highest quality testing in a timely manner.

Microscopy

Confocal laser scanning microscope:

A Zeiss LSM 700 confocal laser scanning microscope (CLSM) is available on a fee-for-service basis. This system features a Zeiss inverted microscope fitted with a laser scanning system with four lasers for advanced imaging of eukaryotic and prokaryotic cells under a variety of conditions.

Primary features of the CLSM:

• Zeiss Axio Observer. Z1 fully automated inverted microscope with XYZ motorized stage with bright field, DIC, and epifluorescence imaging capability
• Objectives: 10x, 20x, 40x (dry), 40x (water), and 63x (oil)
• Four laser lines at 445nm, 488nm, 555nm, and 639nm with two detection channels.
• Zeissís new ZEN software for full featured control of imaging and image analysis.

Additional features:

• A temperature controlled stage for doing incubation experiments.
• Eppendorf TransferMan NK2 micromanipulator system. This fully automated system allows capture and manipulation of single cells for transfer, isolation, or genetic manipulations.

This fully featured microscope allows ready imaging of four different fluors simultaneously, optical sectioning and 3D image reconstruction, photobleaching capacity, time-lapse imaging, tiling of images, as well as other applications consistent with a high grade CSLM. It is also possible to use the system to do time course studies, including time-lapse imaging, directly on the microscope and conduct single cell manipulations.

Image 1: An image of a marine biofilm of a photosynthetic cyanobacteria (red autoflourescence) growing with a background of bacteria (stained blue). Credit Joyce McBeth.

Image 2: Leptothrix cholodnii, a sheathed bacterium; the sheaths are stained red with a maleimide dye specific for sulfhydryl (-SH) groups, while the cells are stained green with a dye for nucleic acids. Credit: Amy Langdon and Emily Fleming.

The Electron Microscope Facility:

The electron microscope facility features (1) a Zeiss 902A transmission electron microscope (TEM) equipped with an EELS system and (2) a Zeiss Supra25 field emission scanning electron microscope (SEM). The TEM is equipped with a goniometer for rotating/tilting sections, the microscope has both digital and film camera systems, and the EELS attachment allows for elemental analysis, removal of inelastic electrons for better resolution and the examination of semi-thick sections (up to 600 nm). The goniometer and the thick section capabilities are excellent tools for producing stereo pair images from moderately thick cell slices, and these can be used reconstructing three-dimensional models of cells. The TEM is supported with two ultramicrotomes, a new RMC PT-X and an older Reichert Ultracut E. Individual scientists are expected to purchase their own diamond knives for cutting thin sections. The field emission SEM provides ultrahigh quality images, and it is equipped with a STEM, energy dispersive X-ray spectrophotometer (EDS) and Robinson diffraction attachments. The STEM (scanning transmission electron microscopy) produces SEM images at the same time as transmitted electron images when viewing certain electron transparent materials (e.g., cells on a grid). EDS is used to examine element composition, especially of biological materials. The Robinson diffraction gives element composition, especially for metal objects. The EM suite has a Denton Vacuum desk-model sputter-coater/evaporator (Desk IV, with Carbon Accessory). This instrument coats grids and SEM specimens with carbon or a variety of metals (e.g., gold, platinium, tungsten). The facility also has a LKB glass knifemaker, a Zeiss SV8 dissecting microscope equipped with ring illumination for examining stubs and grids, a Metler scale, a pH meter, embedding oven, refrigerator with freezer, vacuum pump, and so forth.

Users who wish to operate the system will need to demonstrate proficiency before being allowed to operate unsupervised. There is a $50 per hour beam charge. Users are expected to provide their own supplies (e.g., grids, stubs, stains).

Bigelow Analytical Services

Dr. Stephen Archer, Director

BAS is open to requests for new approaches and analyses. Please email us for more information. Click the links below for more details on BAS services.