Illumination device for spatial and temporal control of morphogen signaling in cell cultures

Abstract

Provided are systems and methods for spatially and temporally controlling light with an illumination device comprising a light source operably connected to a circuit board, one or more light guide plates, one or more optical masks, a controller, and a computer readable medium, comprising instructions that, when executed by the controller, cause the controller to: illuminate a cell or a substrate with light from the light source, and spatially and temporally control illumination of light to the cell or the substrate with one or more illumination parameters, wherein the one or more light guide plates provides uniform illumination of the light. Also provided herein are methods of screening using the system and/or device of the present disclosure.

1 .- 49 . (canceled) 
     
     
         50 . A system for spatially and temporally controlling stem cell or progenitor cell signaling, differentiation, or both, the system comprising:
 (a) a plurality of stem cells or progenitor cells; and   (b) an illumination device positioned to illuminate the plurality of stem cells or progenitor cells, the illumination device comprising:
 (i) a light source that produces light; and 
 (ii) a controller, 
   
       wherein the light source illuminates the plurality of stem cells or progenitor cells with light, 
       wherein the controller spatially and temporally controls illumination of the plurality of stem cells or progenitor cells with one or more illumination patterns comprising one or more illumination parameters, 
       wherein the one or more illumination parameters comprises illumination intensity, and 
       wherein the system spatially and temporally controls stem cell or progenitor cell signaling, differentiation, or both. 
     
     
         51 . The system of  claim 50 , wherein the one or more illumination parameters further comprises one or more of: an illumination duration, an illumination pattern, an illumination wavelength, and any combination thereof. 
     
     
         52 . The system of  claim 50 , wherein the light source comprises one or more light-emitting diodes (LEDs). 
     
     
         53 . The system of  claim 50 , wherein at least one cell of the plurality of stem cells or progenitor cells is genetically engineered to express a light-activatable protein. 
     
     
         54 . The system of  claim 53 , wherein the light source illuminates the plurality of stem cells or progenitor cells with light at a wavelength sufficient to activate the light-activatable protein in the at least one cell. 
     
     
         55 . The system of  claim 53 , wherein an amount of activated light-activatable protein is controlled by the illumination intensity. 
     
     
         56 . The system of  claim 53 , wherein the light-activatable protein is selected from the group consisting of: a cryptochrome, a photoactive yellow protein (PYP) photosensor, a photoreceptor of blue light using flavin adenine dinucleotide (BLUF), a light, oxygen or voltage sensing (LOV) domain, and a phytochrome. 
     
     
         57 . The system of  claim 53 , wherein the light-activatable protein is a light-inducible dimerizer. 
     
     
         58 . The system of  claim 53 , wherein the light-activatable dimerizer is selected from the group consisting of: a CRY2/CIB system, a Phy/Pif system, and a BphP1/PpsR2 system. 
     
     
         59 . The system of  claim 50 , wherein the illumination device is configured to be placed into an incubator. 
     
     
         60 . The system of  claim 50 , wherein the illumination device is wirelessly controllable. 
     
     
         61 . The system of  claim 50 , further comprising one or more light guide plates comprising one or more light guides. 
     
     
         62 . The system of  claim 61 , wherein the one or more light guide plates is configured to provide uniform illumination of the light to at least a subset of the plurality of stem cells or progenitor cells. 
     
     
         63 . The system of  claim 50 , further comprising one or more optical masks. 
     
     
         64 . The system of  claim 63 , wherein the one or more optical masks comprises a photo mask, an intensity mask, or a phase mask. 
     
     
         65 . The system of  claim 50 , wherein at least one cell of the plurality of stem cells or progenitor cells is independently illuminated by the light source. 
     
     
         66 . The system of  claim 50 , wherein the plurality of stem cells or progenitor cells are on a multi-well plate. 
     
     
         67 . The system of  claim 50 , wherein the plurality of stem cells or progenitor cells are on a tissue culture plate. 
     
     
         68 . The system of  claim 50 , further comprising a graphical user interface (GUI) configured to communicate with the controller. 
     
     
         69 . The system of  claim 68 , wherein the GUI is configured to communicate wirelessly with the controller. 
     
     
         70 . The system of  claim 68 , wherein the GUI is configured to allow a user to define the one or more illumination parameters. 
     
     
         71 . The system of  claim 50 , wherein the system is configured to spatially and temporally pattern differentiation of the plurality of stem cells or progenitor cells. 
     
     
         72 . The system of  claim 50 , wherein the illumination intensity is below an amount that causes phototoxicity in the plurality of stem cells or progenitor cells. 
     
     
         73 . The system of  claim 50 , wherein the illumination intensity is from about 0.005 μW/mm 2  to about 20 μW/mm 2 . 
     
     
         74 . The system of  claim 50 , further comprising a pulse generator configured to pulse the light emitted from the light source. 
     
     
         75 . The system of  claim 50 , wherein the one or more illumination patterns is a pulsing pattern, a sinusoidal pattern, a linear pattern, a blinking pattern, or any combination thereof. 
     
     
         76 . The system of  claim 50 , wherein the one or more illumination parameters comprises an illumination duration, and wherein the illumination duration is about 0.5 ms or more. 
     
     
         77 . The system of  claim 50 , wherein the one or more illumination parameters comprises a pulsing frequency, and wherein the pulsing frequency is about 1 Hz or more. 
     
     
         78 . The system of  claim 50 , wherein the one or more illumination parameters comprises an illumination duration, and wherein the illumination duration is about 1 minute or more. 
     
     
         79 . The system of  claim 50 , wherein the light source is configured to illuminate a first subset of the plurality of stem cells or progenitor cells with light at a first wavelength, and to illuminate a second subset of the plurality of stem cells or progenitor cells with light at a second wavelength.