Loading the cassette, Sample volume – ChemoMetec NC-3000 User Manual

Page 24

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2 Advanced cell analysis with the NucleoCounter

®

NC-3000

991-3001 Rev. 1.3

June-2013

8

2.2.2 Loading the Cassette

The Cassette is loaded by fully immersing the tip of the Cassette into the sample

and then gently pressing the white piston, which creates a partial vacuum in the flow

system. This results in the sample being loaded into the flow system. Approximately

60µl is loaded into the flow system.

Figure 3 Loading of the Cassette

The Cassettes to be used with NucleoCounter

®

NC-

3000™ are highly specialized

devices, which hold the key to several of the unique features of the NucleoCounter

®

NC-

3000™ concept. For instance in the Via1-Cassette™ the fluorescent dyes AO

(acridine orange) and DAPI (4',6-diamidino-2-phenylindole) are immobilized inside

the flow channels of the disposable Cassette. W hen the Cassette has been loaded

with approximately 60 µl of sample mixture the dyes are dissolved and mixed with

the sample, staining the cells.

After being placed in the NucleoCounter

®

NC-3000

™, the stained mixture is

automatically transferred to the measurement chamber , where the fluorescent image

is recorded. In a Cassette the measurement volume is as large as 3. 2 µl.

After analysis the sample and dyes are contained inside the Cassette, which can be

safely discarded. This offers a unique and safe sample handling and disposal.

As the Via1-Cassette

™ (and all other cassettes) contains the entire flow system as

well as measurement chamber, neither cleaning nor maintenance of the

NucleoCounter

®

NC-

3000™ instrument itself is needed.

2.2.3 Sample volume

During production, a calibration parameter for all Cassettes is found, which specifies

the precise depth or thickness of the measurement chamber of the device. The

calibration parameter is used by the instrument during analysis and the volume

analyzed is determined by multiplying the imaged area with the depth of the

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