Pico liter well holding device and method of making the same
The present invention broadly comprises a holding device for studying cells comprising at least one cavity adapted to receive cells in a medium consisting essentially of water, the cavity having a substrate and a generally inert wall, wherein the substrate includes a surface for receiving the medium, and wherein the surface includes a multiplicity of pico liter wells and is characterized in that the substrate is substantially translucent and has a refractive index equal to the refractive index of the medium The invention further comprises a method of making the holding device comprising providing a carrier plate, applying an adhesive layer to the carrier plate, depositing a curable substrate on the adhesive layer, applying a second layer of adhesive to the substrate, attaching a wall structure to the second layer of adhesive, forming a multiplicity of pico liter wells in the substrate, curing the substrate, and removing the template.
1. A holding device for studying cells comprising:
at least one cavity adapted to receive a sample of cells in a medium consisting essentially of water, the cavity having a substrate and a generally inert wall, wherein the substrate includes a surface for receiving the medium, and wherein the surface includes a multiplicity of pico liter wells and is characterized in that the substrate is substantially translucent and has a refractive index equal to the refractive index of the medium.
2. The holding device of claim 1 , wherein the medium comprises water and wherein the substrate has a Refractive Index of 1.33.
3. The holding device of claim 1 , wherein the substrate is moldable.
4. The holding device of claim 1 , wherein the substrate is inert.
5. The holding device of claim 1 , wherein the holding device is a carrier plate, and wherein a first adhesive is disposed between the carrier plate and the substrate.
6. The holding device of claim 5 , further including a second adhesive disposed between the generally inert wall and the substrate.
7. The holding device of claim 6 , wherein at least one of the substrate, the first adhesive and the second adhesive are UV-light curable.
8. The holding device of claim 6 , wherein the first adhesive and the second adhesive are acrylic.
9. The holding device of claim 7 , further including a light source transmitting the UV-light through a bottom surface of the at least one cavity.
10. The holding device of claim 9 , wherein the substrate is exposed to the UV-light under vacuum pressure.
11. The holding device of claim 10 , wherein the vacuum pressure is in the range of 0.3-0.5 mmHg.
12. The holding device of claim 9 , wherein the substrate is exposed to the UV-light under inert gas.
13. A method of making a holding device for studying cells, comprising the steps of:
providing a carrier plate;
applying a first adhesive layer to the carrier plate;
depositing a curable substrate on the adhesive layer;
applying a second layer of adhesive to the substrate;
attaching a wall structure to the second layer of adhesive;
forming a multiplicity of pico liter wells in the substrate with a template;
curing the substrate; and
removing the template.
14. The method according to claim 13 , in which the step of curing the substrate includes exposing the substrate to UV-light through the carrier plate.
15. The method according to claim 13 , further comprising repeating the step of curing the substrate after removing the template, wherein the step of curing the substrate after removing the template includes exposing the substrate to UV-light under a vacuum pressure through the carrier plate.
16. The method according to claim 13 , in which the step of forming a multiplicity of pico liter wells in the substrate with a template further comprises embossing a surface of the substrate with a die.
17. A method of making a holding device, comprising the steps of:
providing a layer of curable substrate;
disposing a wall structure on the layer of curable substrate;
embossing a multiplicity of pico liter wells in a surface of the curable substrate; and
curing the substrate.
18. The method according to claim 17 , further comprising the step of disposing a first adhesive between the curable substrate and the wall structure.
19. The method according to claim 17 , in which the step of curing the substrate comprises exposing the substrate to the UV-light through the carrier.
20. A method of making a holding device for studying cells comprising the steps of:
providing a carrier plate having a plurality of cavities surrounded by walls;
depositing a curable substrate in each cavity;
embossing a multiplicity of pico liter wells in the curable substrate with a die; and
curing the substrate.
21. The method according to claim 20 , further comprising the step of applying a first adhesive layer in each cavity before depositing the curable substrate in each cavity.
22. The method according to claim 20 , in which the step of curing the substrate comprises exposing the substrate and the first adhesive to UV-light through the carrier plate when the die is in place.
23. The method according to claim 20 , further comprising the steps of removing the die and exposing the substrate to UV-light under a vacuum or inert gas through the carrier plate.
24. A holding device for studying cells comprising:
a layer of substantially transparent substrate material having a multiplicity of pico liter wells having a refractive index of 1.33; and,
a wall structure attached to the substrate.
25. The holding device of claim 24 , wherein the substrate is UV-light curable.
26. The holding device of claim 24 , further comprising a first adhesive disposed between the wall structure and the substrate.
27. The holding device of claim 24 , further comprising a substantially transparent carrier plate having a plurality of cavities surrounded by walls formed in a first surface of the carrier plate, wherein the layer of substantially transparent substrate material is disposed on the carrier plate.