Lighting systems and methods of using lighting systems for in vitro potency assay for photofrin
View Patent ↗Presently disclosed is a lighting system and methods of using the lighting system for in vitro potency assay for photofrin. The lighting system includes a lamp housing, a first lens, an infrared absorbing filter, an optical filter, and a second lens. The lamp housing includes a lamp and a light-port. In operation, broad spectrum light from the lamp exits the lamp housing by passing through the light-port. The first lens then collimates the broad spectrum light that exits the lamp housing through the light-port. The infrared absorbing filter then passes a first portion of the collimated broad spectrum light to the optical filter and absorbs infrared light of the broad spectrum light. The optical filter then passes a second portion of the collimated broad spectrum light to the second lens. The second lens then disperses the second portion of the collimated light to provide uniform irradiation of a cell culture plate. A method of using the lighting system for studying a photosensitizer is also disclosed.
1. A lighting system comprising:
a lamp housing including a wall, a lamp, and a light-port, wherein the light-port is located within the wall, and wherein broad spectrum light from the lamp exits the lamp housing through the light-port;
a first lens to collimate the broad spectrum light that exits the lamp housing through the light-port;
an infrared absorbing filter to pass a first portion of the collimated broad spectrum light and absorb infrared light of the broad spectrum light that passes through the light-port and that is collimated by the first lens, wherein the first portion of the collimated broad spectrum light comprises a second portion of the collimated broad spectrum light;
an optical filter to pass the second portion of the collimated broad spectrum light after the first portion of the collimated broad spectrum light reaches the optical filter;
a second lens to disperse the second portion of the collimated light that passed through the optical filter, wherein a position of the second lens is adjustable in at least one of a direction parallel to the wall and a direction perpendicular to the wall;
a shelf including a shelf top perpendicular to the wall; and
a lens slider removably attached to the shelf top, wherein the second lens is removably attached to the lens slider, wherein the lens slider includes a first hole for passing the dispersed second portion of the collimated light that passed through the second lens, wherein the shelf includes a second hole for passing the dispersed second portion of the collimated light that passed through the second lens, and wherein at least a portion of the first hole is above or below at least a portion of the second hole.
2. The lighting system of claim 1 , further comprising:
a ring stand;
a first support ring removably attached to the ring stand; and
a base.
3. The lighting system of claim 2 ,
wherein the second lens comprises a first dispersing lens and a second dispersing lens; and
wherein the first support ring holds the first dispersing lens and the second dispersing lens in place.
4. The lighting system of claim 3 ,
wherein the first dispersing lens comprises a first plano convex lens;
wherein the second dispersing lens comprises a second plano convex lens;
wherein the first plano convex lens comprises a first plano side and a first convex side;
wherein the second plano convex lens comprises a second plano side and a second convex side; and
wherein the first convex side is adjacent to the second convex side with a gap between the first convex side and the second convex side.
5. The lighting system of claim 4 , wherein the gap is within the range of 2 millimeters and 4 millimeters, inclusive.
6. The lighting system of claim 5 , wherein the gap is 3 millimeters.
7. The lighting system of claim 1 , wherein the optical filter comprises an infrared blocking filter and a short pass filter.
8. The lighting system of claim 7 , further comprising:
a reflector to reflect the first portion of the collimated broad spectrum light that passes through the infrared absorbing filter.
9. The lighting system of claim 8 , wherein the reflector comprises a dichroic mirror.
10. The lighting system of claim 9 , wherein the dichroic mirror absorbs at least a portion of infrared light that passes through the light-port and the infrared absorbing filter.
11. The lighting system of claim 8 ,
wherein the first portion of the collimated broad spectrum light reflected by the reflector propagates to the infrared blocking filter, and
wherein the second portion of the collimated broad spectrum light that propagates to the infrared blocking filter, as part of the first portion of the collimated broad spectrum light, passes through the infrared blocking filter and then through the short pass filter.
12. The lighting system of claim 8 ,
wherein the first portion of the collimated broad spectrum light reflected by the reflector propagates to the short pass filter, and
wherein the second portion of the collimated broad spectrum light that propagates to the short pass filter, as part of the first portion of the collimated broad spectrum light, passes through the short pass filter and then through the infrared blocking filter.
13. The lighting system of claim 7 ,
wherein the infrared blocking filter absorbs residual infrared light of the first portion of the collimated broad spectrum light, and
wherein the short pass filter filters out light of the first portion of the collimated broad spectrum light of wavelengths greater than 650 nm.
14. A method for studying a photosensitizer, the method comprising:
adding the photosensitizer to a portion of wells on a cell culture plate to form photosensitizer assay wells, the wells comprising carcinoma cells;
incubating the photosensitizer assay wells for a first predetermined time period;
optionally washing the photosensitizer assay wells;
irradiating the photosensitizer assay wells with the lighting system of claim 1 at a predetermined wavelength to form irradiated wells, wherein each well is uniformly irradiated; incubating the irradiated wells for a second predetermined time period; and
determining percent viability of the carcinoma cells contained in the wells.
15. The method according to claim 14 , wherein the step of washing the photosensitizer assay wells is mandatory.
16. The method according to claim 14 , wherein a remaining portion of the wells are control wells.
17. The method according to claim 14 , wherein a portion of the wells contain a reference drug used for comparison.
18. The lighting system of claim 1 , wherein the infrared absorbing filter comprises an infrared absorbing liquid filter.
19. The lighting system of claim 18 , wherein the infrared absorbing liquid filter absorbs between 90 % and 100%, inclusive, of the infrared light of the broad spectrum light that that passes through the light-port.
20. The lighting system of claim 1 ,
wherein the shelf comprises a shelf riser parallel to the wall;
wherein the shelf riser comprises a first parallel adjustment slot; and
wherein the shelf top comprises a first perpendicular adjustment slot.
21. The lighting system of claim 20 , wherein the wall comprises a base wall and a wall of the lamp housing.
22. A method comprising:
using the lighting system of claim 1 to irradiate contents of a cell culture plate.
23. The method of claim 22 , wherein the cell culture plate comprises a photosensitizer.
24. The method according to claim 14 , wherein the photosensitizer is a porphyrin-based anti-neoplastic agent.
25. The method according to claim 24 , wherein the porphyrin-based anti-neoplastic agent is porfimer sodium.
26. The lighting system of claim 1 , wherein the first lens comprises a condenser lens.
27. The lighting system of claim 1 , wherein the first lens comprises a Fresnel lens.
28. The lighting system of claim 1 , wherein the lamp comprises a xenon arc lamp.
29. The lighting system of claim 1 , wherein the optical filter comprises a band-pass filter.
30. The lighting system of claim 1 , wherein the lamp housing is sealed so that the broad spectrum light from the lamp exits the lamp housing only through the light-port.
31. The lighting system of claim 1 , wherein the lamp housing is sealed so that less than 1% of the broad spectrum light from the lamp exits the lamp housing other than through the light-port.
32. The lighting system of claim 1 ,
wherein the lamp housing comprises a top,
wherein the light-port is within the top,
wherein the first lens is within the light-port, and
wherein the infrared absorbing filter, the optical filter, and the second lens are located at positions above the top.
33. The lighting system of claim 1 ,
wherein the lamp housing comprises a bottom,
wherein the light-port is within the bottom,
wherein the first lens is within the light-port, and
wherein the infrared absorbing filter, the optical filter, and the second lens are located at positions below the bottom.
34. The method according to claim 14 , wherein the photosensitizer assay wells are irradiated with light within a range of 400 nm to 650 nm.
35. The method according to claim 14 , wherein the carcinoma cells are A549 human lung carcinoma cells.
36. The method according to claim 14 , wherein the irradiating step is standardized.
37. The method according to claim 14 , wherein the cell culture plate is an opaque black plate.