High-throughput platform for in-vivo sub-cellular screens on vertebrate larvae
High throughput system for in vivo screens on vertebrate larvae. The system includes a source of vertebrate larvae in a liquid medium and loading tube means for aspirating a larva. A detector assembly is provided to differentiate passage of a larva from bubbles and/or debris. An imaging means is provided for both confocal imaging and wide-field fluorescence imaging of the larva. A laser is provided for optical manipulation of the larva.
1. A high-throughput system for in vivo screens comprising:
a source of a specimen organism in a liquid medium;
an automatic syringe pump means to load the specimen organism in the liquid medium into a glass capillary tube;
an automatic optical means to position the specimen organism in the glass capillary tube within a field-of-view of an imaging apparatus for subcellular resolution microscopy through the glass capillary tube; and
a motor and capillary assembly operable to rotate the glass capillary tube, thereby rotating the specimen organism within the field-of-view of the imaging apparatus.
2. The system of claim 1 wherein the specimen organism is a teleost.
3. The system of claim 1 wherein the source is a reservoir containing a plurality of specimen organisms.
4. The system of claim 1 wherein the source is a multiwall plate containing a plurality of specimen organisms.
5. The system of claim 3 wherein the source can be switched between the specimen organisms reservoir and a liquid supply.
6. The system of claim 4 wherein a tube assembly is positioned on the multiwell plate by a position stage.
7. The system of claim 6 wherein the tube assembly includes an injection tube to inject fluid and an aspiration tube to aspirate a specimen organism from the multiwell plate.
8. The system of claim 6 wherein the tube assembly and multiwell plate can be sealed.
9. The system of claim 1 wherein the system includes an automatic detector which has one or more light sources and one or more light detectors arranged in transmission and/or reflection configurations.
10. The system of claim 9 wherein the automatic detector differentiates air bubbles and/or debris from the specimen organisms.
11. The system of claim 1 wherein the tube includes a portion that is a glass or quartz or transparent polymer capillary within the field-of-view of the imaging apparatus.
12. The system of claim 1 wherein the imaging apparatus includes at least one motor for rotating the tube within the field-of-view of the imaging apparatus.
13. The system of claim 1 wherein the imaging apparatus simultaneously images through two objective lenses.
14. The system of claim 1 wherein the imaging apparatus has confocal and/or two-photon and/or wide-field imaging capability.
15. The system of claim 1 wherein the imaging apparatus includes a laser that generates a beam suitable for ablation and/or photoactivation and/or photostimulation.
16. The system of claim 1 wherein the system is used to perform genetic screens.
17. The system of claim 1 wherein the system is used to perform chemical screens.
18. The system of claim 2 wherein the teleost is zebrafish.
19. The system of claim 1 wherein the specimen organism is transgenic.
20. An automated system for in vivo screens on vertebrate larvae comprising:
a source of vertebrate larvae in a liquid medium;
automatic syringe pump means to aspirate a larva in the liquid medium into a glass capillary tube;
an automatic optical detection means to differentiate passage of the larva from bubbles or debris in the glass capillary tube;
an imaging apparatus configured to image the larva in the glass capillary tube; and
a motor and capillary assembly operable to rotate the glass capillary tube, thereby rotating the larva within a field of view of the imaging apparatus.
21. The system of claim 20 wherein the larva is a zebrafish larva.
22. The system of claim 20 wherein the imaging apparatus includes a confocal microscope.
23. The system of claim 20 wherein the imaging apparatus includes a brightfield microscope.