Institute of Neurobiology

Investigators’ research laboratories:

Dr. S. Treistman’s lab space consists of 1000 sq. ft. of newly renovated space in the Institute of Neurobiology. It houses electrophysiology setups, molecular biology lab with fume hood, sink, etc., cell culture lab including cell culture hood and incubators, several computer work stations, and desk space for lab personnel. Office space, shared instrument room, tissue culture room, cold-room, dark-room, and dishwashing and autoclaving facility are on the same floor.

Dr. J. Rosenthal’s lab consists of 800 sq. ft. of newly renovated space in the Institute of Neurobiology. It houses a student office, space for 2 electrophysiology rigs, a fumehood, space for 8 molecular biology/biochemistry workstations, a small machine shop, a walk-in coldroom, and a patch-clamp room. Dr. Rosenthal has a separate office of ~100 sq. ft.  A Xenopus laevis colony is housed in special tanks in the Institute’s animal facility.

Dr. B. Marie has two neighboring laboratories of 710 ft². One lab is solely dedicated to electrophysiology and contains a sink, two dissection set up and two electrophysiology set up. This lab can accommodate 2 researchers. The second lab is designed to carry out imaging, molecular biology and genetics. It contains a dark room for imaging. It has 2 refrigerator/freezers, molecular biology apparatus, a sink, microscopes and 2 apparatus to anesthetize and cross flies. It is designed for 7 people. The P.I. has an office of approximately 220 ft2 close to the laboratory.

Dr. M. Sosa’s lab at the Institute of Neurobiology consists of  (380 ft2) with a refrigerator, separate freezer, incubator, sink, 2 dissection tables, 1 vibration-isolation table, electrophysiology recording setup, analytical balance, microcentrifuge, pH meter, fume hood, cabinets for glassware and chemicals, 2 dissection microscopes, 1 epifluorescence microscope, 1 standard light microscope, 4 computers, 3 printers, 1 fax machine, storage closet, and shelf space. The PI also has bench space assigned in the Molecular Neurobiology Core Facility at the basement of the Institute of Neurobiology (1,245 ft2), consisting of 2 large and 1 small room fully equipped for molecular biology.

Dr. M. Miller has a laboratory on the first floor of the Institute of Neurobiology of approx. 550 ft2. This lab has a refrigerator, a separate freezer, two incubators, two sinks, two dissection tables, a vibration-isolation table, two electrophysiology recording setups, a set-up for recording heartbeat, balances, a microcentrifuge, a pH meter, cabinets for glassware and chemicals, 2 dissection microscopes, an epifluorescence microscope, three computers, storage closets, and shelf space.

Dr. Yudowski’s lab consists of ~1000 sq. ft. of newly renovated lab space with an adjacent state-of-the-art imaging facility in the Institute of Neurobiology. The imaging space includes a dedicated area for live cell microscopy housing a TIRF microscope with 4 laser lines and a EMCCD camera. The lab is specifically designed to conduct cellular and molecular biology experiments. It houses cell and tissue culture incubators, a dedicated molecular biology station and standard lab equipment.  A specially designed spinning disk microscope for water samples is located in the same building.

Institute of Neurobiology Core Facilities:

The Molecular Neurobiology Core consists of ~ 450 sq. ft. in the Institute of Neurobiology. It has 6 molecular work stations, a -70°C freezer, 5 water baths, 5 air incubators, 1 bacterial incubator, 1 high-speed centrifuge, 1 benchtop refrigerated centrifuge, 3 microcentrifuges, 1 shaker, 1 Millipore ultrafiltration system, 1 ice machine, 4 refrigerators, 2 freezers, 3 thermal cycling PCR machines, 1 Real-time PCR machine, 7 pipet sets, 1 balance, 1 pH meter, 1 autoclave, 4 gel electrophoresis systems, 1 gel transfer/blot unit, a PC equipped with gel documentation and analysis system/software and thermal printing device, a Macintosh computer with sequence analysis software and an HP Laserjet printer, and separate RNA, PCR and radioactive compound rooms.

The Neurogenetics Core consists of ~ 450 sq. ft that contains a facility to make Drosophila food, 2 incubators used to maintain 750 different stocks and 2 apparatus to anesthetize and cross flies.

The Microscopy Core contains a Zeiss Pascal LSM5 Confocal microscope and a Live Imaging System that combines Zeiss microscopy with spinning disk technology.  A TIRF microscope will soon be available. This microscope will allow our scientists to visualize in real time the movement of receptor  molecules in the membrane of cells, including neurons, with a resolution that has not been possible before.

 

Institute of Neurobiology Animal Facitities

Blue crab (Callinectes sapidus) specimens are trapped locally and kept in natural light conditions on the ground floor of the Institute. Water is also brought from the collection sites.  Maintenance of the tanks is carried out by the Institute of Neurobiology Animal Care Facility personnel.

Freshwater prawns are kept in rack-mounted 15 gallon individual tanks, linked through a common filtration system, and on 3 cylindrical 350 gallon plastic enclosures on the Animal Care Facilities at the Institute of Neurobiology or the Main Campus Building. Maintenance of the tanks is carried out by the personnel of the Animal Care Facilities

A colony of 30-60 Xenopus are maintained in a Xenopus Aquatic Housing System from Allentown Aquaneering. This system has particulate, biological and carbon filters, constant flow, automatic water exchange, pH and ammonia monitoring, and temperature control. Frogs are fed three times a week by a technician and monitored daily. Each week oocytes are harvested from 2 animals. Each animal is operated on a maximum of 6 times, with at least 12 weeks between operations.

 

Institute of Neurobiology Visiting Scientist Facilities

The present remodeling project includes living quarters (400 sq ft), including cooking facilities and a private bathroom for a visiting scientist. In addition to the lab benches (Molecular Neurobiology Core) and instrumentation (Microscopy and Neurogenetics Cores) that are presently available, remodeling plans also include dedicated lab space (450 sq ft) for visiting scientists.

 

Major Equipment:

 

Dr. Treistman’s lab: 6 Olympus microscopes with epifluorescence and Hoffman optics, 2 dissecting microscopes, 1 video camera, 1 air table, 6 Sutter micromanipulators, 6 patch clamp amplifiers, 2 oscilloscopes, 1 chart recorder, 2 4-channel AC amplifiers, 1 Dagan Instruments single/double electrode voltage clamp, David Kopf vertical micropipette puller, Sutter Instruments P 97 micropipette puller, Picospritzer pressure injection system.

Dr. Rosenthal’s lab: There are three functioning electrophysiology rigs. The first is a 2 microelectrode voltage clamp (Geneclamp 500B from Axon Instruments). The second is a cut-open oocyte clamp (CA-1B High Performance Oocyte Clamp from Dagan Instruments). Both of these rigs have home built peltier driven temperature controllers for the bath. The last rig is a patch clamp (Axopatch 200B from Axon Instruments) equipped with a rapid solution changer (RSC-200 from Biologic) and a line temperature controller (TC-324 B from Warner Instruments). For all rigs, D/A and A/D signal processing uses Innovated Integrations boards that samples up to 333 nS/point. GPATCH, the software used to drive the system, was kindly provided by Dr. Francisco Bezanilla. For pipette fabrication we use a Sutter P-97 puller and an MF-200 microforge (WPI).  Oocytes are injected with a Nanoliter 2000 microinjector (WPI). In support of the elctrophysiology rigs (e.g. making chambers) there is a machine shop in my lab with a mini milling machine and lathe as well as equipment for building electronics. All basic items for molecular biology are available. These include multiple agarose and acrylamide gel rigs, a FIGE gel set-up, power supplies, two PTC-200 thermocylers from MJ Research, a gel documentations system, a Nanodrop spectrophotometer, stationary and shaking 37ºC incubators, a Beckman L8-60M Ultrcentrifuge and a Beckman J2-21 High Speed Centrifuge, an autoclave, refrigerators, and a -80ºC and 2 -20ºC freezers.  All DNA sequences are sent to the DNA Sequencing Core at the UPR-Rio Piedras. Radioactive and fluorescent imaging of gels uses a Typhoon 9200 Phoshphor/fluorescence imager located in the Molecular Core facility of the Institute of Neurobiology which is adjacent to my lab. Specific items for Pichia pastoris include a stationary and a rotating 28ºC incubator and a French Press to disrupt the yeast.

Dr. B. Marie Lab 1: 2 Stereo Microscopes with fiber-optic light sources, 2 Nikon FN1 microscopes (1 with DIC optics and epifluorescence; 1 with DIC only), 2 air tables, 2 sutter translators, 2 PCLAMP10 CNS software for windows, 2 HP computers, 2 Axopatch 200B-2 Microelectrode Amplifiers, 2 Huxley-style micropositioners, 2 Digidata 1440A low-noise data acquisition system, 2 stimulus isolators, 2 Master-8 progammable pulse stimulators AMPI, 2 MP285 automated Sutter micromanipulators, 2 Prior micromanipulators, 1 Sutter P-97 micropipette puller, 1 Narishige microelectrode polisher, 1 Sutter Beveller BV-10.  Lab 2: 2 Stereo Microscopes, 2 light sources, 1 Nikon 80i microscope with DIC optics and epifluorescence, 1 Nikon SMZ800 microscope with GFP fluorescence. Molecular biology: 2 apparatus to run DNA/ RNA gels; 2 apparatus to run and blot Protein gels, water bath, incubator, Gel imaging system,  hot blocks, thermocycler, refrigerated centrifuge, room temperature centrifuge, 3 shakers, 2 apparatus to anesthetize and cross flies.

Dr. María Sosa’s lab has an electrophysiological recording setup with a Gateway 1.7 GHz Pentium IV computer, AcqKnowledge data acquisition software and an HP DeskJet Printer. The laboratory also has a Dell Optiplex GX260 2.0 GHz Pentium IV, a Gateway EL2-500SE 2.0 GHz Pentium IV and a Gateway E-6100 3.0 GHz Pentium IV computer, which share a HP LaserJet 1200 series and a HP 2110 all-in-one printer. Dr. Sosa’s office has a Gateway 2.2 GHz Pentium IV computer with an HP Laserjet 6P and a Kodak 8660 Thermal Printer.