Systems and methods for preparing and analyzing low volume analyte array elements

Abstract

Serial and parallel dispensing tools that can deliver defined and controlled volumes of fluid to generate multi-element arrays of sample material on a substrate surface are provided. The substrates surfaces can be flat or geometrically altered to include wells of receiving material. Also provided are tools that allow the parallel development of a sample array. To this end, the tool can be understood as an assembly of vesicle elements, or pins, where each of the pins can include a narrow interior chamber suitable for holding nanoliter volumes of fluid. Each of the pins can fit inside a housing that forms an interior chamber. The interior chamber can be connected to a pressure source that will control the pressure within the interior chamber to regulate the flow of fluid within the interior chamber of the pins. The prepared sample arrays can then be passed to a plate assembly that disposes the sample arrays for analysis by mass spectrometry.

We claim:  
     
         1 . A dispensing apparatus for dispensing nanovolumes of fluid in chemical or biological procedures onto the surface of a substrate, comprising 
 a housing having a plurality of sides and a bottom portion having formed therein a plurality of apertures, said walls and bottom portion of said housing defining an interior volume,    one or more fluid transmitting vesicles, mounted within said apertures, having a nanovolume sized fluid holding chamber for holding nanovolumes of fluid, said fluid holding chamber being disposed in fluid communication with said interior volume of said housing, and    dispensing means in communication with said interior volume of said housing for selectively dispensing nanovoluments of fluid from said nanovolume sized fluid transmitting vesicles when the fluid is loaded with said fluid holding chambers of said vesicles, whereby said dispensing means dispenses nanovolumes of the fluid onto the surface of the substrate when the apparatus is disposed over and in registration with the substrate.    
     
     
         2 . The apparatus of    claim 1   , wherein each said fluid transmitting vesicle has an open proximal end and a distal tip portion that extends beyond said housing bottom portion when mounted within said apertures, said open proximal end disposing said fluid holding chamber in fluid communication with said interior volume when mounted with the apertures.  
     
     
         3 . The apparatus of    claim 1   , wherein said plurality of fluid transmitting vesicles are removably and replaceably mounted within said apertures of said housing.  
     
     
         4 . The apparatus of    claim 1   , wherein said plurality of fluid transmitting vesicles include a glue seal for fixedly mounting said vesicles within said housing.  
     
     
         5 . The apparatus of    claim 1   , wherein said fluid holding chamber includes a narrow bore dimensionally adapted for being filled with the fluid through capillary action.  
     
     
         6 . The apparatus of    claim 1   , wherein each said fluid holding chamber of said plurality of fluid transmitting vesicles are sized to fill substantially completely with the fluid through capillary action.  
     
     
         7 . The apparatus of    claim 1   , wherein said plurality of fluid transmitting vesicles comprise an array of fluid delivering needles.  
     
     
         8 . The apparatus of    claim 7   , wherein said fluid delivering needles are formed of metal.  
     
     
         9 . The apparatus of    claim 7   , wherein said fluid delivering needles are formed of glass.  
     
     
         10 . The apparatus of    claim 7   , wherein said fluid delivering needles are formed of silica.  
     
     
         11 . The apparatus of    claim 7   , wherein said fluid delivering needles are formed of polymeric material.  
     
     
         12 . The apparatus of    claim 1   , wherein the number of said plurality of fluid transmitting vesicles is less than or equal to the number of wells of a multi-well substrate.  
     
     
         13 . The apparatus of    claim 1   , wherein said housing further includes a top portion, an further comprising mechanical biasing means of mechanically biasing said plurality of fluid transmitting vesicles into sealing contact with said housing bottom portion.  
     
     
         14 . The apparatus of    claim 13   , wherein each said fluid transmitting vesicle has a proximal end portion that includes a flange, and further comprising a sealer element disposed between the flange and an inner surface of the housing bottom portion for forming a seal between the interior volume and an external environment.  
     
     
         15 . The apparatus of    claim 14   , wherein said mechanical biasing means includes a plurality of spring elements each of which are coupled at one end to said proximal end of each said plurality of fluid transmitting vesicles, and at another end to an inner surface of said housing top portion, said spring element applying a mechanical biasing force to said vesicle proximal end to form said seal.  
     
     
         16 . The apparatus of    claim 1   , wherein said housing further includes a top portion, and further comprising securing means for securing said housing top portion to said housing bottom portion.  
     
     
         17 . The apparatus of    claim 16   , wherein said securing means comprises a plurality of fastner-receiving apertures formed within one of said top and bottom portions or said housing, and a plurality of fastners for mounting within said apertures for securing together said housing top and bottom portions.  
     
     
         18 . The apparatus of    claim 1   , wherein said dispensing mens comprises a pressure source fluidly coupled to said interior volume of said housing for disposing said interior volume at a selected pressure condition.  
     
     
         19 . The apparatus of    claim 18   , wherein said fluid transmitting vesicles are filled through capillary action, and wherein said dispensing means further comprises means for varying said pressure source to dispose said interior volume of said housing at varying pressure conditions, said means for varying disposing said interior volume at a selected pressure condition sufficient to offset said capillary action to fill the fluid holding chamber of each vesicle to a predetermined height corresponding to a predetermined fluid amount.  
     
     
         20 . The apparatus of    claim 1     9 , wherein said means for varying further comprises fluid selection means for selectively discharging a selected nanovolume fluid amount from said chamber of each said vesicle.  
     
     
         21 . The apparatus of    claim 1   , wherein said fluid transmitting vesicle has a proximal end that opens onto said interior volume of sid housing, and wherein said fluid holding chamber of said vesicles are sized to substantially completely fill with the fluid through capillary action without forming a meniscus at said proximal open end.  
     
     
         22 . The apparatus of    claim 1   , wherein said dispensing means comprises fluid selection means for selectively varying the amount of fluid dispensed from said fluid holding chamber of each vesicle.  
     
     
         23 . The apparatus according to    claim 1   , having plural vesicles, wherein a first portion of said plural vesicles include fluid holding chambers of a first size and a second portion including fluid holding chambers of a second size, whereby plural fluid volumes can be dispensed.  
     
     
         24 . The apparatus of    claim 22   , wherein said fluid selection means comprises a pressure source coupled to said housing and in communications with said interior volume for disposing said interior volume at a selected pressure condition, and 
 adjustment means coupled to said pressure source for varying said pressure within said interior volume of said housing to apply a positive pressure in said fluid chamber of each said fluid transmitting vesicle to vary the amount of fluid dispensed therefrom.    
     
     
         25 . A fluid dispensing apparatus for dispensing a fluid in chemical or biological procedures into one or more wells of a multi-well substrate, comprising 
 a housing having a plurality of sides and a bottom portion having formed therein a plurality of apertures, said walls and bottom portion defining an interior volume,    a plurality of fluid transmitting vesicles, mounted within said apertures having a fluid holding chamber disposed in communication with said interior volume of said housing,    a fluid selection and dispensing means in communication with said interior volume of said housing for variably selecting an amount of the fluid loaded with said fluid holding chambers of said vesicles to be dispensed from a single set of plurality of fluid transmitting vesicles, and    whereby said dispensing means dispenses a selected amount of the fluid into the wells of the multi-well substrate when the apparatus is disposed over and in registration with the substrate.    
     
     
         26 . The fluid dispensing apparatus of    claim 25   , wherein said fluid selection and dispensing means is adapted to select various amounts of fluid to be dispensed from said single set of vesicles.  
     
     
         27 . The fluid dispensing apparatus of    claim 25   , wherein said fluid selection and dispensing means comprises a pressure source fluidly coupled to said interior volume of said housing for disposing said interior volume at a selected pressure condition.  
     
     
         28 . The fluid dispensing apparatus of    claim 27   , further compromising means for varying the pressure within the interior volume of the housing to select the amount of fluid to dispense from said fluid transmitting vesicles.  
     
     
         29 . The fluid dispensing apparatus of    claim 27   , wherein said fluid transmitting vesicles are filled with the fluid through capillary action, and further comprising means for varying said pressure source to dispose said interior volume of said housing at varying pressure conditions, said means for varying disposing said interior volume at a pressure condition sufficient to offset said capillary action to fill the fluid holding chamber of each vesicle to a predetermined height corresponding to a predetermined fluid amount.  
     
     
         30 . The fluid dispensing apparatus of    claim 25   , wherein said fluid selection means comprises 
 a pressure source coupled to said housing and in communication with said interior volume for disposing said interior volume at a selected pressure condition, and    adjustment means coupled to said pressure source for varying said pressure within said interior volume of said housing to apply a positive pressure in said fluid chamber of each said fluid transmitting vesicle to vary the amount of fluid dispensed therefrom.    
     
     
         31 . A fluid dispensing apparatus for dispensing fluid in chemical or biological procedures into one or more wells of a multi-well substrate, said apparatus comprising 
 a housing having a plurality of sides and top and bottom portions of said bottom portion having formed therein a plurality of apertures, said walls and top and bottom portions of said housing defining an interior volume,    a plurality of fluid transmitting vesicles, mounted within said apertures having a fluid holding chamber sized to hold nanovolumes of the fluid, said fluid holding chamber being disposed in fluid communication with said volume of said housing and    mechanical biasing means for mechanically biasing said plurality of said transmitting vesicles into sealing contact with said housing bottom portion.    
     
     
         32 . The fluid dispensing apparatus of    claim 31   , wherein each said fluid transmitting vesicle has a proximal end portion that includes a flange, and further comprising a sealer element disposed between the flange and an inner surface of the housing bottom portion for forming a pressure and fluid seal between the internal and external environment.  
     
     
         33 . The fluid dispensing apparatus of    claim 31   , wherein said mechanical biasing means includes a plurality of spring elements each of which are coupled at one end to said means includes a plurality of spring elements each of which are coupled at one end to said proximal end of said fluid transmitting vesicle, and at another end to an inner surface of said housing top portion, said spring elements applying a mechanical biasing force to said vesicle proximal end to form said fluid and pressure seal.  
     
     
         34 . The fluid dispensing apparatus of    claim 31   , further comprising securing means for securing said housing top portion to said housing bottom portion.  
     
     
         35 . The fluid dispensing apparatus of    claim 34   , wherein said securing means comprises a plurality of fastener-receiving apertures formed within one of said top and bottom portions of said housing, and a plurality of fasteners for mounting within said apertures for securing said housing top and bottom portions together.  
     
     
         36 . The fluid dispensing apparatus of    claim 31   , further comprising dispensing means in communication with said interior volume of said housing for selectively dispensing the fluid from said fluid transmitting vesicles when the fluid is loaded within said fluid holding chambers of said vesicles, whereby said dispensing means dispenses the fluid into the wells of the multi-well substrate when the apparatus is disposed over an in registration with the substrate.  
     
     
         37 . The fluid dispensing apparatus of    claim 36   , wherein said dispensing means comprises a pressure source fluidly coupled to said interior volume of said housing for disposing said interior volume at a selected pressure condition.  
     
     
         38 . The fluid dispensing apparatus of    claim 31   , wherein said plurality of fluid transmitting vesicles are removably and replaceably mounted within said apertures of said housing.  
     
     
         39 . The fluid dispensing apparatus of    claim 31   , wherein said plurality of fluid transmitting vesicles comprises an array of fluid delivering needles.  
     
     
         40 . The fluid dispensing apparatus of    claim 36   , wherein said fluid transmitting vesicles are filled with the fluid through capillary action, and wherein said dispensing means further comprises means for varying said pressure source to dispose said interior volume of said housing at varying pressure conditions, said means for varying disposing said interior volumes at a selected pressure condition sufficient to offset said capillary action to fill the fluid holding chamber of each vesicle to a predetermined height corresponding to a predetermined fluid amount.  
     
     
         41 . The fluid dispensing apparatus of    claim 36   , wherein said dispensing means comprises fluid selection means for selectively varying the amount of fluid dispensed from said fluid holding chamber of each vesicle.  
     
     
         42 . The fluid dispensing apparatus of    claim 31   , further comprising a pressure source coupled to the housing and in communication with the interior volume for disposing the interior volume at a selected pressure condition, and 
 adjustment means coupled to the pressure source for varying the pressure within the interior volume of the housing to apply a positive pressure to the fluid chamber of each the fluid transmitting vesicle to vary the amount of fluid dispensed therefrom.