Biomedical Engineering Reference
In-Depth Information
4.5.9 Use of PubChem
The value of PubChem cannot be underestimated. We have found it remarkably useful
to follow the activity of molecules in UNMCMD screens across the MLPCN. This
tracking capability identifies bona fide activity in screens on other targets. The library
has also been characterized with respect to compounds that aggregate, fluoresce, or
are otherwise reactive species.
4.6 FUTURE OPPORTUNITIES
As we see it, flow cytometry in a small-molecule discovery environment will benefit
from ongoing development of software and hardware. The instrumentation leader for
the UNMCMD, Bruce Edwards now has an NIH-funded R01 for improved sample
handling and automation (1R01HG005066-01 Advancing High Throughput Flow
Cytometry). The work is coordinated with the overall automation of the HyperCyt
platform to improve sample detection, transition to 1536-well sampling, and integrate
a direct sampling system for the flow cytometer.
4.6.1 Bubble Technology: Optical Time Stamping System for Robust
Indexing of HyperCyt Flow Cytometry Data
Edwards has been developing an air bubble detection system to supplement our
current sample indexing protocols. As a sample-separating bubble passes through
the point of analysis of the flow cytometer, a flash of light is produced due to laser
light scattering by the air-liquid interface. Both flow cytometry data and bubble
data are being recorded in the same FCS format, facilitating integration of the two
data streams in the analysis software. We are now in process of developing an optical
fiber interface that will extend the range of instruments amenable to bubble data
collection.
4.6.2 Adaptation of HyperCyt for Processing of 1536-Well Plates
We are developing 1536-well plate processing capabilities so that a single HyperCyt
system can operate four sampling probes per 1536-well plate to deliver 384-well
segments simultaneously to four Accuri C6 flow cytometers. Accuri Corporation
has released an upgraded version of CFlow software called “CFlow for the
HyperCyt System.” This software provides an interface by which the C6 cytometer
can be remotely controlled over a network via TCP/IP. Also, the software allows
collection of up to 96 million events per round of data acquisition, a significant
advance over the original 1 million event limit. We have developed TCP/IP software
to communicate with the CFlow interface so that a laptop PC controls all four flow
cytometers in parallel via an Ethernet network in addition to its control of the
HyperCyt autosampler. Multiplex samples in 1536-well plates have been delivered
in 11min.
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