Table 7. Bleed through test.
| Dye | Illumination | Cy5 filter | Qdot filter | Cy3 filter |
|---|---|---|---|---|
| Qdot 525 | PE4000 405nm | |||
| PE4000 435nm | ||||
| PE4000 460nm | + | |||
| PE2 425nm | ||||
| Sola | AUTO | |||
| Qdot 565 | PE4000 405nm | |||
| PE4000 435nm | ||||
| PE4000 460nm | ||||
| PE2 425nm | ||||
| Sola | +++ (**) | |||
| Qdot 585 | PE4000 405nm | |||
| PE4000 435nm | ||||
| PE4000 460nm | ||||
| PE2 425nm | ||||
| Sola | ++ | |||
| Qdot 605 | PE4000 405nm | |||
| PE4000 435nm | ||||
| PE4000 460nm | ||||
| PE2 425nm | ||||
| Sola | ++ | |||
| Qdot 625 | PE4000 405nm | |||
| PE4000 435nm | ||||
| PE4000 460nm | ||||
| PE2 425nm | ||||
| Sola | + & AUTO (***) | |||
| Qdot 655 | PE4000 405nm | |||
| PE4000 435nm | ||||
| PE4000 460nm | ||||
| PE2 425nm | ||||
| Sola | + | AUTO | ||
| Cy5 | PE4000 635nm | |||
| Sola | ||||
| PE2 615nm | (*) | |||
| PE2 425nm | ||||
| Cy3 | PE 4000 535nm | |||
| PE4000 405nm | ++ | |||
| PE4000 435nm | ++ | |||
| PE4000 460nm | ++ | |||
| Sola | ++ | |||
| PE2 535nm | ||||
| PE2 425nm | ++ |
Serial sections stained with the indicated dye (column 1) were illuminated with the indicated light (column 2) and of the observation of specific fluorescence (+/++/+++) and autofluorescence (AUTO) going through each filter block (Cy5; Qdot & Cy3 filter blocks) was recorded.
+/++/+++ reflect the intensity of the specific fluorescence; AUTO reflects the presence of noticeable autofluorescence.
grey cells: no visible fluorescence; green cells: expected specific fluorescence, strong unless indicated; pink cells: bleed through (i.e. observation of fluorescence in a set up where fluorescence is not desirable).
Excitation wave lengths that would not normally be used for a dye are indicated by a red cell.
(*) more autofluorescence was observed with PE2 615nm than with Sola & PE4000 635nm
(**) see Fig 10;
(***) see image in S2 Fig