Interactive transparent individual cells biochip processor

• US 20050014201A1
• Filed: 10/25/2001
• Published: 01/20/2005
• Est. Priority Date: 10/25/2001
• Status: Abandoned Application

• Current Assignee: BAR ILAN University
• Original Assignee: BAR ILAN University
• Inventors: Deuthsch, Mordechai
• Application Number:
  US10/492,531
• Publication Number:
  US 20050014201A1
• Time in Patent Office: Days
• US Class Current: 435/7.200
• CPC Class Codes: B01J 2219/00317 Microwell devices, i.e. having large numbers of wells
• CPC Class Codes: B01J 2219/00702 Processes involving means for analysing and characterising the products
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: C12M 23/12 Well or multiwell plates C12M25/04 takes precedence
• CPC Class Codes: C12M 25/08 electrically charged
• CPC Class Codes: C12M 41/46 of cellular or enzymatic activity or functionality, e.g. cell viability
• CPC Class Codes: C40B 60/14 for creating libraries
• CPC Class Codes: G01N 15/1433 using image recognition
• CPC Class Codes: G01N 15/1484 microstructural devices
• CPC Class Codes: G01N 2015/1006 for cytology
• CPC Class Codes: G01N 2015/1472 with colour
• CPC Class Codes: G01N 2015/1497 Particle shape
• CPC Class Codes: G01N 2035/00158 Elements containing microarrays, i.e. "biochip"
• CPC Class Codes: G01N 2035/0429 Sample carriers adapted for special purposes
• CPC Class Codes: G01N 33/5005 involving human or animal cells immunoassay G01N33/56966; immunoassays of protozoa G01N33/56905; protozoa in screening assays C12Q1/025
• CPC Class Codes: G02B 21/34 Microscope slides, e.g. mounting specimens on microscope slides

Abstract

An Interactive Transparent Individual Cells Biochip Processor (ITICBP) device is described, which is useful for assessing a single, individual living cell at identifiable location or assessing group of cells each at identifiable location.

Claims

1-75. -75. (Canceled).

76. A device for holding a plurality of living cells, comprising:

(a) a plurality of addressable, at least partially optically transparent, wells;

(b) a cell-mover, configured to move individual living cells into and/or out of said wells; and

(c) a reagent dispenser, configured to dispense at least one reagent into said wells.

77. The device of claim 76, wherein each said well is configured to hold a plurality of said living cells.

78. The device of claim 76, wherein each said well is configured to hold a single said living cell.

79. The device of claim 76, further comprising at least one assessor instrument, said assessor instrument configured to assess at least one cell property selected from the group consisting of cell morphology, cell activity, cell physiology, cell metabolism, cell affinity, cell viability, cell changes that occur as a result of presence or absence of contact with other cells and/or specific biologically-active materials.

80. The device of claim 76, further comprising at least one image acquiring device selected from the group consisting of chromatic image acquiring device, fluoroscopy and electron microscope image acquiring device.

81. The device of claim 76, further comprising a computer device configured to perform at least one function from the group functions consisting of control functions of the device, record the status of each individual living cell, record the device status and analyze acquired data.

82. The device of claim 76, wherein the said wells are substantially hexagonal.

83. The device of claim 76, wherein said wells are of a shape selected from the group consisting of circular, square, rectangular and triangular.

84. The device of claim 76, further comprising walls separating a said well from a neighboring said well.

85. The device of claim 84, wherein said walls are substantially optically transparent.

86. The device of claim 76, wherein said wells are made of a material selected from the group consisting of glass, substantially transparent polymer, polycarbonate, nylon, laminated PTFE, non-laminated PTFE, cellulose acetate, glass filter and cellulose ester.

87. The device of claim 76, wherein a material from which said wells are made is porous and/or perforated so as to allow a fluid to flow through said material.

88. The device of claim 76, further comprising at least one pore or perforation at the bottom of said wells so as to allow a fluid to flow through said wells.

89. The device of claim 76, further comprising a plurality of spacers configured to support a cover plate above said wells.

90. The device of claim 89, further comprising a cover plate provided with at least one electrode, said at least one electrode configured to induce an electrical field so as to alter the position of a said living cell found within a said well.

91. The device of claim 76, further comprising at least one electrode, said at least one electrode configured to induce an electrical field so as to alter the position of a said living cell found within a said well.

92. The device of claim 76, further comprising at least one biosensor electrode, said biosensor electrode configured to detect materials released by a said living cell found within a said well.

93. The device of claim 92, wherein said biosensor electrode is configured to convert a chemical reaction response to a signal that is processable by transduction.

94. The device of claim 92, wherein said biosensor electrode is configured to produce a variable signal, the variation in said variable signal related to a quantity of a detected change, said detected change relating to an amount of a material, an ion, a radical, electrons, heat, mass or light released by a said living cell found within a said well.

95. The device of claim 92, wherein said biosensor electrode is coated with a biologically active material, said biologically active material configured to selectively react with said released material, and wherein said biosensor electrode is configured to produce a variable signal, the variation in said variable signal related to a quantity of said released material reacting with said biologically active material within a said well.

96. The device of claim 95, wherein said biologically active material is selected from the group consisting of enzymes, antibodies, immunogens, antigens, ligands, receptors, nucleic acids, proteins, peptides and receptors.

97. The device of claim 76, further comprising microtunnels, said microtunnels configured to allow communication of fluids between two said wells.

98. A chip-device for holding living cells, the device comprising a carrier having a plurality of individually addressable wells disposed on an upper surface each well configured to hold at least one living cell, and a lower surface, the device characterized in having at least one feature from amongst the four features:

(a) at least one of said wells is defined by an intersection of at least two channels on said upper surface;

(b) said carrier is provided with a transport channel configured to transport fluids from one location of said carrier to another location of said carrier;

(c) having at least one well microelectrode associated with at least one of said wells; and

(d) wherein said wells are juxtaposed.

99. The device of claim 98, wherein said wells are configured to hold no more than one living cell of a certain type.

100. The device of claim 98, wherein the dimensions of said wells are less than about 200 microns.

101. The device of claim 98, wherein the dimensions of said wells are less than about 50 microns.

102. The device of claim 98, wherein the dimensions of said wells are less than about 25 microns.

103. The device of claim 98, wherein the dimensions of said wells are less than about 10 microns.

104. The device of claim 98, further comprising channels for transporting fluids from said upper surface to said lower surface through said carrier.

105. The device of claim 104, wherein said channels are pores in said wells, said pores being of a size so as to prevent passage of a said living cell.

106. The device of claim 98, further comprising a biologically active material bound to a said well through a photolysable linker.

107. The device of claim 106, wherein said biologically active material is selected from the group consisting of a chemical material, a biological material, a drug, an enzyme, an antibody, an immunogene, an antigen, a ligand, a receptor, a nucleic acid, a protein and a peptide.

108. The device of claim 98, wherein said carrier is substantially transparent.

109. The device of claim 98, wherein said upper surface is substantially transparent and said lower surface is substantially not transparent.

110. The device of claim 98, wherein said lower surface is substantially transparent and said upper surface is substantially not transparent.

111. The device of claim 98, wherein at least one of said wells is defined by an intersection of at least two channels on said upper surface.

112. The device of claim 111, wherein said at least two intersecting channels are transport channels configured to transport fluids from one location of said carrier to another location of said carrier.

113. The device of claim 98, further comprising at least one transport channel on said upper surface configured to transport fluids from one location of said carrier to another location of said carrier.

114. The device of claim 113, wherein said at least one transport channel is configured to transport fluids from one said well to another said well.

115. The device of claim 113, wherein said at least one transport channel separates at least two groups of said wells.

116. The device of claim 98, wherein said wells are enclosures of dimensions such that substantially an entire cell of a certain type is containable within a said enclosure, each said enclosure having an opening at said upper surface, said opening defined by a first cross section of a size allowing passage of a cell of a certain size.

117. The device of claim 116, wherein the volume of said enclosure is in the order of 10⁻

11 liter.

118. The device of claim 116, wherein the area of said first cross section is less than about 25000 micron².

119. The device of claim 116, wherein the area of said first cross section is less than about 1500 micron².

120. The device of claim 116, wherein the area of said first cross section is less than about 350 micron².

121. The device of claim 116, wherein said upper surface is substantially transparent so as to allow observation of a cell contained within a said enclosure therethrough and said lower surface is substantially not transparent.

122. The device of claim 116, wherein said lower surface is substantially transparent so as to allow observation of a cell contained within a said enclosure therethrough and said upper surface is substantially not transparent.

123. The device of claim 116, wherein the dimensions of a said enclosure are such as to contain no more than one said cell of a certain size at any one time.

124. The device of claim 116, wherein said enclosures are substantially parallel walled and a said first cross section and a second cross section of said enclosure parallel thereto are substantially identical.

125. The device of claim 124, wherein said second cross section is substantially of a shape selected from the group consisting of circular, hexagonal, triangular, square and rectangular.

126. The device of claim 116, wherein said enclosures are substantially parallel walled and a said first cross section and a second cross section of said enclosure parallel thereto are substantially not identical.

127. The device of claim 126, wherein said second cross section is substantially of a shape selected from the group consisting of circular, hexagonal, triangular, square and rectangular.

128. The device of claim 116, wherein a distance between rims of two openings of two adjacent enclosures is less than 20% of a smallest dimension of said first cross section.

129. The device of claim 116, wherein a distance between rims of two openings of two adjacent enclosures is less than 10% of a smallest dimension of said first cross section.

130. The device of claim 116, wherein a distance between rims of two openings of two adjacent enclosures is less than 6% of a smallest dimension of said first cross section.

131. The device of claim 98, wherein said wells are juxtaposed.

132. The device of claim 131, having at least seven said wells, wherein the total interstitial area between said seven wells is less than or equal to about 0.35 the sum of the areas of said seven wells.

133. The device of claim 131, having at least seven said wells, wherein the total interstitial area between said seven wells is less than or equal to about 0.25 the sum of the areas of said seven wells.

134. The device of claim 131, having at least seven said wells, wherein the total interstitial area between said seven wells is less than or equal to about 0.15 the sum of the areas of said seven wells.

135. The device of claim 131, having at least seven said wells, wherein the total interstitial area between said seven wells is less than or equal to about 0.10 the sum of the areas of said seven wells.

136. The device of claim 131, having at least seven said wells, wherein the total interstitial area between said seven wells is less than or equal to about 0.06 the sum of the areas of said seven wells.

137. The device of claim 131, wherein said wells are hexagonally packed.

138. The device of claim 131, wherein a rim of a said well is substantially knife-edged.

139. The device of claim 131, further comprising at least one protuberance protruding from said upper surface between two adjacent wells.

140. The device of claim 139, further comprising a cover plate configured to rest on said protuberance so as to define an aperture between said two adjacent wells, the size of said aperture such that a said cell of the certain size cannot pass therethrough.

141. The device of claim 131, further comprising at least two protuberances protruding from said upper surface between two adjacent wells, the space between said at least two protuberances being such that a said cell of the certain size cannot pass therebetween.

142. The device of claim 141, further comprising a cover plate configured to rest on said protuberances so as to define an aperture between said two adjacent wells, the size of said aperture such that a said cell of the certain size cannot pass therethrough.

143. The device of claim 98, further comprising at least one well microelectrode associated with a said well.

144. The device of claim 143, comprising a plurality of independently activatable well microelectrodes, each one of said plurality of well microelectrodes associated with a different said well

145. The device of claim 143, wherein said well microelectrode is configured to act through said carrier.

146. The device of claim 143, wherein said well microelectrode is embedded within said carrier.

147. The device of claim 143, wherein said well microelectrode is found within said associated well.

148. The device of claim 143, wherein said well microelectrode is transparent.

149. The device of claim 143, wherein said well microelectrode is configured to apply an electrical stimulus to a cell found within said associated well.

150. The device of claim 143, wherein said well microelectrode is configured to apply a repulsive force to a cell found within said associated well.

151. The device of claim 150, wherein said well microelectrode is positioned so that a said applied repulsive force is directed away from said associated well.

152. The device of claim 143, wherein said well microelectrode is configured to apply an attractive force to a cell found within said associated well.

153. The device of claim 152, wherein said well microelectrode is positioned so that a said applied attractive force is directed towards said associated well.

154. The device of claim 143, wherein associated with at least one said well are at least two well microelectrodes.

155. The device of claim 154, wherein two of said at least two well microelectrodes are electrically isolated from each other.

156. The device of claim 154, wherein said two well microelectrodes are configured to simultaneously each apply a force on a cell found within said associated well so as to generate torque on said cell.

157. The device of claim 143, wherein said well microelectrode is a biosensor electrode.

158. The device of claim 157, wherein said biosensor electrode is configured to detect a release of light by a cell found within said associated well, a release of heat by a cell found within said associated well or a change of mass of a cell found within said well.

159. The device of claim 157, wherein said biosensor electrode is configured to detect a material released by a cell found within said associated well.

160. The device of claim 159, wherein said released material is selected from the group consisting of ions, radicals, electrons and molecules.

161. The device of claim 159, wherein said biosensor electrode is configured to quantify said released material.

162. The device of claim 159, wherein associated with a said well are at least three biosensor electrodes.

163. The device of claim 159, wherein said at least three biosensor electrodes are configured to act together as a transducer to convert said detection of said released material to an electrical signal.

164. The device of claim 159, wherein said biosensor electrode comprises a biologically active material chosen to specifically react with said released material.

165. The device of claim 164, wherein said biologically active material is selected from the group consisting of a chemical material, a biological material, a drug, an enzyme, an antibody, an immunogene, an antigen, a ligand, a receptor, a nucleic acid, a protein and a peptide.

166. The device of claim 98, further comprising at least one wall protruding from said upper surface, said at least one wall circumscribing at least one area of said upper surface where the points of the top edge of said wall define a plane.

167. The device of claim 166, wherein said wall is continuous.

168. The device of claim 166, further comprising a cover plate configured to rest on said top edge of said at least one wall so as to define at least one closed volume including more than one said well.

169. The device of claim 166, wherein said at least one wall circumscribes more than one area of said upper surface.

170. The device of claim 169, wherein said at least one wall is continuous.

171. The device of claim 169, further comprising a cover plate configured to rest on said top edge of said wall so as to define at least two closed volumes, said at least two closed volumes each including more than one said well.

172. The device of claim 98, further comprising a cover slip, said cover slip and said carrier configured so as to allow said cover slip to removeably rest above said upper surface substantially in parallel to said upper surface.

173. The device of claim 172, wherein said configuration of said carrier includes at least one wall protruding from said upper surface, allowing said cover slip to rest thereupon substantially in parallel to said upper surface.

174. The device of claim 172, wherein said configuration of said carrier includes at least three protrusions protruding from said upper surface, said protrusions having substantially the same height, allowing said cover slip to rest thereupon substantially in parallel to said upper surface.

175. The device of claim 172, wherein said cover slip and said carrier are configured so that there exists a limited number of correct cover slip positions wherein said cover slip is substantially oriented in a specific position when resting above said upper surface.

176. The device of claim 175, wherein said limited number is selected from the group consisting of one, two, four and six.

177. The device of claim 172, wherein said cover slip further comprises at least one cover slip microelectrode.

178. The device of claim 177, wherein said cover slip microelectrode is transparent.

179. The device of claim 177, wherein said cover slip microelectrode is configured to apply an attractive force on a cell found between said cover slip and said upper surface when said cover slip rests above said upper surface.

180. The device of claim 177, wherein said at least one cover slip microelectrode is configured to apply a repulsive force on a cell found between said cover slip and said upper surface when said cover slip rests above said upper surface.

181. The device of claim 176, wherein said cover slip microelectrode is associated with a specific well when resting above said carrier.

182. The device of claim 181, wherein said cover slip microelectrode is configured to apply an attractive force on a cell found in a said associated well so as to extract said cell from said well when said cover slip rests above said upper surface.

183. The device of claim 181, wherein said cover slip microelectrode is configured to apply a repulsive force on a cell found in the vicinity of said cover slip microelectrode so as to cause said cell to approach said associated well when said cover slip rests above said upper surface.

184. The device of claim 181, wherein also associated with said associated well is at least one well microelectrode.

185. The device of claim 184, wherein said at least one well microelectrode and said at least one cover slip microelectrode are configured to simultaneously each apply a force to a cell so as to generate torque on a cell found in the vicinity of said associated well.

186. The device of claim 98, further comprising a flow generator, configured to generate a flow of fluid substantially parallel to said upper surface.

187. A method for rotating a living cell, comprising:

(a) providing at least two microelectrodes; and

(b) simultaneously applying a force from said at least two microelectrodes on the cell so as to generate torque on the cell.

188. A method for agitating a living cell, comprising repetitively:

(a) producing a first force having a first direction using a first microelectrode; and

(b) producing a second force having a second direction using a second microelectrode wherein said second direction is substantially opposite said first direction.

189. The method of claim 188, wherein said first force and said second force are repulsive.

190. The method of claim 188, wherein said first force and said second force are attractive.

191. The method of claim 188, wherein said first microelectrode and said second microelectrode apply respective said forces to a cell found inside an enclosure.

192. The method of claim 191, wherein said enclosure has a volume in the order of 1000 micron³.

193. A method for studying living cells, comprising:

(a) providing a group of cells;

(b) isolating cells from said group of cells each of said cells in an individual well, each said well being individually addressable;

(c) lysing at least one said isolated cell so that said lysates remain in said individual well; and

(d) examining said lysates.

194. The method of claim 193, wherein said lysis is performed using a method selected from the group consisting of thermal lysis, chemical lysis, sonification, application of an enzyme and application of detergent.

195. A method for selecting cells, comprising:

a. providing a group of cells;

b. isolating cells from said group of cells each of said cells in an individual well, each said well provided with a force generator;

c. examining each isolated cell; and

d. selecting isolated cells fulfilling at least one criterion.

196. The method of claim 195, further comprising:

e. separating said selected cells from said group of cells.

197. The method of claim 196, wherein said separating comprises:

(i) generating a force using said force generator in wells containing said selected cells so as to distance said selected cells from a respective said well; and

(ii) applying a flow of fluids to carry away said selected cells.