HIGH-CONTENT IMAGING OF MICROFLUIDIC DEVICES
The present invention is related to high-content microscopy imaging of microfluidic cell culture systems. A method of high-content microfluidic device microscopy is contemplated, along with related statistical analysis and microfluidic device adaptors.
1 - 110 . (canceled)
111 . A method of applying lipid nanoparticles (LNPs) to cells, comprising a) providing i) a plurality of lipid nanoparticles (LNPs) ii) a microfluidic device comprising a first microchannel seeded with cells of a first cell type, and b) introducing said LNPs into said microfluidic device.
112 . The method of claim 111 , wherein said LNPs comprise nucleic acid sequences.
113 . The method of claim 112 , wherein said nucleic acid sequences are selected from the group consisting of ribonucleic acid (RNA), messenger ribonucleic acid (mRNA), and deoxyribonucleic acid (DNA).
114 . The method of claim 112 , wherein said nucleic acid sequences are silencing ribonucleic acids (RNA) selected from the group consisting of small interfering RNA (siRNA) and RNA interference (RNAi).
115 . The method of claim 112 , wherein said nucleic acid sequences encode a green fluorescent protein (GFP) transgene.
116 . The method of claim 111 , wherein said cells of a first cell type are attached to a membrane in said first microchannel.
117 . The method of claim 111 , wherein said cells of a first cell type are hepatocytes.
118 . The method of claim 111 , wherein said microfluidic device comprises first and second microchannels.
119 . The method of claim 118 , wherein said LNPs are introduced into said first or second microchannel.
120 . The method of claim 111 , wherein said LNPs are introduced in step b) by flowing said LNPs into said microfluidic device.
121 . The method of claim 116 , wherein said microfluidic device is further seeded with cells of a second cell type.
122 . The method of claim 111 , further comprising c) detecting delivery of LNPs to said cells of a first cell type.
123 . The method of claim 112 , further comprising c) detecting the effect of said delivery of nucleic acids on said cells of a first cell type.
124 . The method of claim 112 , further comprising c) detecting cellular phenotype changes following nucleic acid delivery to said cells of a first cell type.
125 . A method of applying lipid nanoparticles (LNP) to cells in a microfluidic device, comprising a) providing i) a plurality of lipid nanoparticles (LNP) comprising nucleic acid sequences; ii) a microfluidic device comprising a microchannel seeded with cells of a first cell type, and b) flowing said LNPs into said microfluidic device for delivering said nucleic acid sequences to said cells.
126 . The method of claim 125 , wherein said nucleic acid sequences are selected from the group consisting of ribonucleic acid (RNA), messenger ribonucleic acid (mRNA), and deoxyribonucleic acid (DNA).
127 . The method of claim 125 , wherein said nucleic acid sequences are silencing ribonucleic acids (RNA) selected from the group consisting of small interfering RNA (siRNA) and RNA interference (RNAi).
128 . The method of claim 125 , wherein said nucleic acid sequences encode a green fluorescent protein (GFP) transgene.
129 . The method of claim 125 , wherein said cells of a first cell type are attached to a membrane in said microchannel.
130 . The method of claim 125 , wherein said cells of a first cell type are hepatocytes.
131 . The method of claim 130 , wherein said hepatocytes are selected from the group consisting of human, monkey, rat and mouse hepatocytes.
132 . The method of claim 125 , wherein said microfluidic device comprises first and second microchannels.
133 . The method of claim 132 , wherein said LNPs are introduced into said first or second microchannel.
134 . The method of claim 133 , wherein said microfluidic device is further seeded with cells of a second cell type.
135 . The method of claim 125 , further comprising c) detecting delivery of LNPs to said cells of a first cell type.
136 . The method of claim 125 , further comprising c) detecting the effect of said delivery of nucleic acids on said cells of a first cell type.
137 . The method of claim 125 , further comprising c) detecting cellular phenotype changes following nucleic acid delivery to said cells of a first cell type.
138 . A method of applying Adeno-Associated Virus (AAV) vectors to cells, comprising a) providing i) a plurality of Adeno-Associated Virus (AAV) vectors ii) a microfluidic device comprising a first microchannel seeded with cells of a first cell type, and b) introducing said AAV vectors into said microfluidic device.
139 . The method of claim 138 , wherein said AAV vectors comprise nucleic acid sequences.
140 . The method of claim 139 , wherein said nucleic acid sequences encode a green fluorescent protein (GFP) transgene.
141 . The method of claim 138 , wherein said cells of a first cell type are attached to a membrane in said first microchannel.
142 . The method of claim 138 , wherein said cells of a first cell type are hepatocytes.
143 . The method of claim 138 , wherein said microfluidic device comprises first and second microchannels.
144 . The method of claim 143 , wherein said AAV vectors are introduced into said first or second microchannel.
145 . The method of claim 138 , wherein said AAV vectors are introduced in step b) by flowing said AAV vectors into said microfluidic device.
146 . The method of claim 143 , wherein said microfluidic device is further seeded with cells of a second cell type.
147 . The method of claim 138 , further comprising c) detecting delivery of said AAV vectors to said cells of a first cell type.
148 . The method of claim 138 , further comprising c) detecting the effect of said delivery of AAV vectors on said cells of a first cell type.
149 . The method of claim 138 , further comprising c) detecting cellular phenotype changes following delivery of said AAV vectors to said cells of a first cell type.
150 . A method of delivering nucleic acid sequences to cells in a microfluidic device, comprising a) providing i) a plurality of Adeno-Associated Virus (AAV) vectors comprising nucleic acid sequences; ii) a microfluidic device comprising a microfluidic channel seeded with cells of a first cell type, and b) flowing said AAV vectors into said microchannel for delivering said sequences to said cells.
151 . The method of claim 150 , wherein said (AAV) vectors are selected from the group of serotypes consisting of AAV2, AAV8, and AAV9.
152 . The method of claim 150 , wherein said cells of a first cell type are attached to a membrane in said microchannel.
153 . The method of claim 150 , wherein said cells of a first cell type are hepatocytes.
154 . The method of claim 153 , wherein said hepatocytes are selected from the group consisting of human, monkey, rat and mouse hepatocytes.
155 . The method of claim 150 , wherein said microfluidic device comprises first and second microchannels.
156 . The method of claim 155 , wherein said AAV vectors are introduced into said first or second microchannel.
157 . The method of claim 150 , wherein said microfluidic device is further seeded with cells of a second cell type.
158 . The method of claim 150 , further comprising c) detecting delivery of said AAV vectors to said cells of a first cell type.
159 . The method of claim 150 , further comprising c) detecting the effect of said delivery of AAV vectors on said cells of a first cell type.
160 . The method of claim 150 , further comprising c) detecting cellular phenotype changes following delivery of said AAV vectors to said cells of a first cell type.