This disclosure demonstrates an approach that translates synthetic DNA codes to spatial codes registered in nanoliter microchambers for multiplexed measurement of nearly any type of molecular targets (e.g., miRNAs, mRNAs, intracellular and surface proteins) in single cells.

Provided herein is a method for detecting the presence of clade variants in the COVID-19 virus in a human sample and/or an environmental sample. Samples are processed to obtain total RNA. The RNA is used as a template in a combined reverse transcription and amplification reaction to obtain fluorescent COVID-19 virus amplicons. These amplicons are hybridized on a microarray with nucleic acid probes having sequences that discriminate among the various clade variants. The microarray is imaged to detect the clade variant and each clade variant is distinguished from others by generating an intensity distribution profile from the image, which is unique to each of the clade variants.

Provided herein is a fluid delivery method for permeabilizing a biological sample. The method includes delivering the fluid to a first substrate and/or a second substrate. At least one of the first substrate and the second substrate includes a spacer. The method further includes assembling, subsequent to the delivering, a chamber comprising the first substrate, the second substrate, the biological sample, and the spacer. The spacer may be disposed between the first substrate and second substrate. The spacer may be configured to maintain the fluid within the chamber and maintain a separation distance between the first substrate and the second substrate. The spacer may be positioned to at least partially surround an area on the first substrate on which the biological sample is disposed and/or at least partially surround the array disposed on the second substrate.

Provided herein is a fluid delivery method for permeabilizing a biological sample. The method includes delivering the fluid to a first substrate and/or a second substrate. At least one of the first substrate and the second substrate includes a spacer. The method further includes assembling, subsequent to the delivering, a chamber comprising the first substrate, the second substrate, the biological sample, and the spacer. The spacer may be disposed between the first substrate and second substrate. The spacer may be configured to maintain the fluid within the chamber and maintain a separation distance between the first substrate and the second substrate. The spacer may be positioned to at least partially surround an area on the first substrate on which the biological sample is disposed and/or at least partially surround the array disposed on the second substrate.

Provided herein are methods of releasing an extended capture probe from a substrate and uses of the same.

Provided herein are methods of releasing an extended capture probe from a substrate and uses of the same.

Provided herein are methods of releasing an extended capture probe from a substrate and uses of the same.

Disclosed herein are compositions and methods for determining a presence or abundance of an analyte in a biological sample. The methods disclosed herein include: (a) providing a biological sample on a substrate comprising a plurality of capture probes, wherein a capture probe of the plurality of capture probes comprises a capture domain; (b) releasing the analyte from the biological sample; (c) affixing a stretching moiety to the analyte; (d) hybridizing the analyte to the capture domain of the capture probe; (e) applying a stretching force to the stretching moiety, thereby elongating the analyte hybridized to the capture domain; and (f) generating an extended capture probe using the analyte as a template.

Disclosed herein are compositions and methods for determining a presence or abundance of an analyte in a biological sample. The methods disclosed herein include: (a) providing a biological sample on a substrate comprising a plurality of capture probes, wherein a capture probe of the plurality of capture probes comprises a capture domain; (b) releasing the analyte from the biological sample; (c) affixing a stretching moiety to the analyte; (d) hybridizing the analyte to the capture domain of the capture probe; (e) applying a stretching force to the stretching moiety, thereby elongating the analyte hybridized to the capture domain; and (f) generating an extended capture probe using the analyte as a template.

Disclosed herein are compositions and methods for determining a presence or abundance of an analyte in a biological sample. The methods disclosed herein include: (a) providing a biological sample on a substrate comprising a plurality of capture probes, wherein a capture probe of the plurality of capture probes comprises a capture domain; (b) releasing the analyte from the biological sample; (c) affixing a stretching moiety to the analyte; (d) hybridizing the analyte to the capture domain of the capture probe; (e) applying a stretching force to the stretching moiety, thereby elongating the analyte hybridized to the capture domain; and (f) generating an extended capture probe using the analyte as a template.