Supported by CCR Office of Science and Technology Resources (OSTR)


Project Development
Image Analysis

HiTIF High-Throughput Microscopes

The vast majority of the assays run in the HiTIF are performed either in a 96- or 384-well clear bottom imaging plates. The HiTIF possesses dedicated instruments for automated liquid handling (Fluorescent stains, Immuno-Fluorescence (IF), RNA FISH, or DNA FISH staining), image acquisition and image analysis:

Yokogawa CV8000 High-throughput spinning disk confocal microscope

The Yokogawa CV8000 is HiTIF second high-throughput imaging system. The CV8000 feature set is similar to the CV7000.

When compared to the latter, the CV8000 has a newer generation of sCMOS cameras (QE 82%, 2048 X 2048 pixels), and a 40X water objective with higher NA (1.15). The CV8000 is not currently integrated with a robotic arm.

Yokogawa CV7000 microscope with integrated robotic arm and plate hotels for automated plate feeding

The Yokogawa CV7000S high-throughput imaging system is a fully automated dual Nipkow spinning microscope equipped with 4 laser lines (405, 488, 561, 640 nm), IR- and image-based autofocus, 2 large sCMOS cameras (2560 X 2048 pixels) and 6 objectives (4X, 10X, 20X LWD, 20X, 40X and 60X water).

The supported plate formats are 6-, 12-, 24-, 48-, 96-, 384-well and sample slides. The CV7000 can image in confocal, epifluorescence, Digital Interference Contrast (DIC) and brightfield mode. The CV7000S includes an integrated environmental chamber which maintains constant temperature, CO2 and humidity for time-lapse, live- cell experiments for up to 72 hrs. In addition the CV7000S is equipped with an integrated liquid dispenser that can transfer reagents from a 96- or 384- well plate into an imaging assay plate during live-cell experiments. Finally, the CV7000 is integrated with a robotic arm, plate hotels, and scheduling software to run automated HTI experiments on up to 42 fixed plates.

Additional Instrumentation

PerkineElmer Janus Automated Liquid Handler

The Janus is equipped with an 8-channel arm for flexible dispensing from reservoirs, conical tubes, microcentrifuge tubes, and plates into 96- or 384-well plates.

In addition, the Janus includes multi-dispense head technology for fast dispensing of liquids into 96- and 384-well plates and two multi-tower plate stackers for a total capacity of 200 assay plates. The Janus is typically used for oligo RNA library resuspension.
Beckman ECHO525 Acoustic Liquid Handler

The ECHO525 uses focused sound waves to transfer liquids from any well of a source plate to any well of a destination plate in 25 nl increments. Source plates must be ECHO compatible (6-well reservoir, 384-well polypropylene, 384-well low dead volume).

Destination plates can be in any format, provided they respect SBS standards and are not taller than 16 mm. The ECHO525 is used in library reformatting/management, siRNA or sgRNA oligos transfection optimization, and automated reverse transfection screening. In addition, the ECHO525 can be used for setup of qRT-PCR reactions, or synthetic biology assembly reactions in 384-well plates.
Biotek EL406 Plate washer/Dispenser

The Biotek EL406 Plate washer can be programmed for automated and semi-automated IF and DNA FISH protocols. It is equipped for with a washer module, a peristaltic pump based cassette dispenser and with two syringe pumps dispensers. If desired, plates can be processed in batch thanks to scheduling software and a Biostack plate stacker integrated with the EL406.

BlueCatBio Bluewasher/Dispenser

The Bluewasher is a tip-less plate washer. It uses centrifugation to discard liquid from 96- or 384-well plates. The Bluwasher software enables controlling centrifugal rotation speed to either minimize residual volumes of liquids in the well post centrifugation or, conversely, minimizing the disruption of cells monolayers in the well.

In addition, the Bluwasher can dispense up to 7 different fluids in 5 microliters increments using a fully automated switching valve. The Bluewasher is used for PFA cell fixation, RNA FISH, and DNA FISH protocols.
PerkinElmer Columbus

Columbus is an image storage and analysis server. HiTIF is equipped with a Columbus servers with 80 TB of dedicated hard-drive space for images, experimental metadata and results.

The Columbus server is equipped with multi-core processors for fast image analysis with the Acapella software; and a database for the storage and organization of images and HCI results. Images, analysis pipelines and results on the Columbus servers can be accessed by HiTIF users from any computer on the NIH network through a web browser interface.