Acoustic Radiation for Ejecting and Monitoring Pathogenic Fluids

Abstract

The invention relates to methods and devices that use focused radiation to handle and/or monitor pathogenic fluids. In particular, a method is provided for dispensing one or more droplets of a fluid containing a pathogen. The method involves providing the pathogen-containing fluid in a reservoir and applying focused radiation to the pathogen-containing fluid in the reservoir in a manner effective to eject a droplet of the fluid therefrom. Often, a pathogen-impermeable enclosure is used.

1 . A method for sealing a fluid in a pathogen-impermeable enclosure, comprising:
 (a) providing the fluid in a reservoir;   (b) positioning a substrate so that a designated site on a surface thereof is in droplet-receiving relationship with respect to the reservoir;   (c) applying focused radiation to the fluid in the reservoir in a manner effective to eject a droplet of the fluid therefrom onto the substrate surface at the designated site; and   (d) sealing the fluid droplet at the designated site in the pathogen-impermeable enclosure.   
     
     
         2 . The method of  claim 1 , wherein the fluid is a bodily fluid. 
     
     
         3 . The method of  claim 1 , wherein the focused radiation is focused acoustic radiation. 
     
     
         4 . The method of  claim 3 , further comprising, before (d), (d°) depositing a pathogen-containing fluid at the designated site. 
     
     
         5 . The method of  claim 4 , wherein the pathogenic fluid is a bodily fluid. 
     
     
         6 . The method of  claim 4 , wherein the pathogen-containing fluid is comprised of a discrete pathogenic particle in a carrier fluid. 
     
     
         7 . The method of  claim 6 , wherein the pathogen is a virus. 
     
     
         8 . The method of  claim 6 , wherein the pathogen is bacterial. 
     
     
         9 . The method of  claim 4 , wherein the pathogen is a toxin. 
     
     
         10 . The method of  claim 4 , wherein (d°) is carried out after (c). 
     
     
         11 . The method of  claim 4 , wherein (d°) is carried out during (c). 
     
     
         12 . The method of  claim 4 , wherein (d°) is carried out before (c). 
     
     
         13 . The method of  claim 3 , wherein (b) comprises (b′) opening the pathogen-impermeable enclosure so as to expose the designated site within the enclosure to the reservoir. 
     
     
         14 . The method of  claim 10 , wherein the pathogen-impermeable enclosure comprises a cover and the substrate, and (b′) comprises positioning the cover to expose the designated site within the enclosure to the reservoir. 
     
     
         15 . The method of  claim 11 , wherein (d) comprises positioning the cover in sealing contact with the substrate. 
     
     
         16 . The method of  claim 15 , further comprising, after (d), (e) subjecting the pathogen impermeable enclosure to temperatures associated with PCR thermal cycling. 
     
     
         17 . The method of  claim 3 , wherein steps (b) and (c) are repeated such that a plurality of droplets is deposited on the substrate surface at different designated sites, and wherein the fluid droplets at each designated site are sealed in a pathogen-impermeable enclosure. 
     
     
         18 . The method of  claim 17 , wherein the different designated sites form an array of sites. 
     
     
         19 . The method of  claim 17 , wherein (d) comprises individually isolating each of the fluid droplets in a compartment of the pathogen-impermeable enclosure. 
     
     
         20 . The method of  claim 3 , wherein (a) comprises providing a plurality of reservoirs each containing a different fluid, and steps (b) and (c) are repeated such that a droplet from each reservoir is deposited on the substrate surface. 
     
     
         21 . The method of  claim 20 , wherein the droplets are deposited on the substrate surface at the same designated site. 
     
     
         22 . The method of  claim 20 , wherein the droplets are deposited on the substrate surface at different designated sites. 
     
     
         23 . A method for monitoring for a change in the amount and/or concentration of a pathogen in a pathogenic fluid, comprising:
 (a) providing a pathogen-impermeable enclosure enclosing the pathogenic fluid, wherein the pathogenic fluid comprises a pathogen and a carrier fluid; and   (b) acoustically monitoring for a change in the amount and/or concentration of the pathogen enclosed in the pathogen-impermeable enclosure.   
     
     
         24 . The method of  claim 23 , wherein the change is an increase in pathogen content. 
     
     
         25 . The method of  claim 23 , wherein the change is a decrease in pathogen content. 
     
     
         26 . The method of  claim 23 , further comprising, before (b), (a′) introducing additional material into the reservoir. 
     
     
         27 . The method of  claim 23 , further comprising, after (a) and before (b), (a′) subjecting the enclosure to a temperature change. 
     
     
         28 . The method of  claim 27 , wherein (a′) comprises raising the temperature of the enclosure. 
     
     
         29 . The method of  claim 27 , wherein (a′) comprises subjecting the enclosure to a temperature selected to facilitate an increase in the amount and/or concentration of the pathogen in the enclosure. 
     
     
         30 . The method of  claim 27 , wherein (a′) comprises subjecting the enclosure to a temperature selected to facilitate a decrease in the amount and/or concentration of the pathogen in the enclosure. 
     
     
         31 . A method for selecting a localized volume in a pathogenic fluid for removal from a reservoir, comprising:
 (a) providing the pathogen-containing fluid in a reservoir comprised of plurality of particles and a carrier fluid; and   (b) acoustically locating the localized volume within the pathogen-containing fluid.   
     
     
         32 . The method of  claim 31 , further comprising, after (b), (c) removing the localized volume from the fluid reservoir. 
     
     
         33 . The method of  claim 32 , wherein (c) is carried out by applying focused radiation to eject a droplet containing the localized volume from the reservoir. 
     
     
         34 . The method of  claim 33 , wherein the focused radiation applied is focused acoustic radiation. 
     
     
         35 . The method of  claim 31 , wherein at least some of the particles are pathogenic particles. 
     
     
         36 . The method of  claim 31 , wherein at least some of the particles are nonpathogenic particles. 
     
     
         37 . The method of  claim 31 , wherein the plurality comprises at least one pathogenic particle and at least one nonpathogenic particle. 
     
     
         38 . The method of  claim 31 , wherein the localized volume contains at least one particle. 
     
     
         39 . The method of  claim 37 , wherein the localized volume contains a single particle. 
     
     
         40 . The method of  claim 31 , wherein the localized volume contains no particle. 
     
     
         41 . The method of  claim 31 , wherein the localized volume contains a pathogen. 
     
     
         42 . The method of  claim 31 , wherein the localized volume contains no pathogen. 
     
     
         43 . A method for detecting for an interaction between a fluid and a compound, wherein the fluid and/or compound are pathogenic, comprising:
 (a) providing a reservoir containing the fluid;   (b) depositing the compound onto a designated site on a surface of a substrate;   (c) positioning the substrate so that the designated site is in droplet-receiving relationship with respect to the reservoir;   (d) applying focused radiation to the fluid in the reservoir in a manner effective to eject a droplet of the fluid therefrom onto the substrate surface at the designated site;   (e) sealing the fluid droplet and the compound at the designated site in a pathogen-impermeable enclosure; and   (f) detecting for an interaction between the fluid and the compound.   
     
     
         44 . The method of  claim 43 , wherein the compound is pathogenic. 
     
     
         45 . The method of  claim 43 , wherein the fluid is pathogenic. 
     
     
         46 . The method of  claim 43 , wherein the interaction is acoustically detected. 
     
     
         47 . The method of  claim 43 , wherein the interaction is optically detected. 
     
     
         48 . The method of  claim 43 , wherein the interaction is fluorescently detected. 
     
     
         49 . The method of  claim 43 , wherein the interaction is magnetically detected. 
     
     
         50 . The method of  claim 43 , wherein the interaction is electrically detected. 
     
     
         51 . The method of  claim 43 , further comprising, after (e), (e′) subjecting the pathogen-impermeable enclosure to temperatures associated with PCR thermal cycling. 
     
     
         52 . A method for transferring one or more droplets of a pathogen-containing fluid, comprising:
 (a) providing the pathogen-containing fluid in a reservoir which lies within a sealed enclosure; and   (b) applying focused acoustic radiation to the pathogen-containing fluid in the reservoir in a manner effective to transfer a droplet of the fluid onto a surface within the sealed enclosure by means of an acoustic radiation generator external to the sealed enclosure, wherein the acoustic radiation generation is coupled to the reservoir by means of an acoustic coupling fluid.   
     
     
         53 . The method of  claim 52 , further comprising the steps of (c) providing a second pathogen-containing fluid in a second reservoir within the sealed enclosure, (d) moving the acoustic radiation generator so that it is able to direct focused acoustic radiation to the second pathogen-containing fluid, and (e) applying focused acoustic radiation to the second pathogen-containing fluid within the sealed enclosure by means of the acoustic radiation generator in a manner effective to transfer a droplet of the fluid onto a surface within the sealed enclosure, wherein the acoustic radiation generator is coupled to the second reservoir by means of an acoustic coupling fluid.