Biomedical Engineering Reference
In-Depth Information
(e) wash slides (0.2
×
SSC): two cycles, source 3, waste 1 at 25
◦
C, flow for 10 s, hold
for 20 s
(f) wash slides (0.1
×
SSC): two cycles, source 4, waste 1 at 25
◦
C, flow for 10 s, hold
for 20 s.
14 Assemble up to six hybridization units (two slides per unit): insert rubber O-rings in the
covers and put the slides on the black bottom plate. Make sure the slides are in the
proper orientation with the printed side up.
15 Introduce the unit into the hybstation, press unit down with one hand while tightening
the screw with the other.
16 Insert plugs into the sample ports and the waste tubes into the corresponding wash
bottles.
17 On the touch screen subsequently press: start a run, from floppy, CGH.hyb, load, the
positions of the slides you want to use, start, continue (the hybstation starts to warm
up the slides).
18 When the hybstation is ready (visible on screen by indication of the module you have to
start) apply hyb mix:
(a) press Probe to add the hyb mix for the selected slide
(b) check if a mark on the screen appears
(c) take the plug out and inject the hyb mix by pipetting it slowly into the port using a
200
μ
l pipette
(d) press the Finished control (check mark) and replace the plug
(e) repeat this for the next slide
(f) press the Finished control for the selected slide
(g) press the Finished control for the module
(h) repeat this for the selected module(s).
19 Take slides out after 38 h and put them in 0.01
×
SSC.
20 Place each slide in a 50ml tube and centrifuge at 200
g
for 3min to dry.
21 Immediately scan slides in a microarray scanner.
22 Cleaning the hybstation:
u
23 Reassemble all used hybridization units with dummy slides and introduce them into the
hybstation.
24 Insert plugs into all sample ports and place all tubes in a bottle of water.
25 On the touch screen subsequently press: maintenance, Machine Cleaning Cycle, the
positions of the slides you used, continue.
26 When cleaning is finished, take out the hybridization units, rinse with water (never use
ethanol) especially the sample port and dry the unit with the air pistol.
