Methods and compositions are provided for isolating protoplasts from plants and other multicellular, cell-wall containing organisms with high efficiency.

The present invention is a technology for stably maintaining a specimen (for example, exfoliative cells of a human body) contained in a vial so that the specimen can be safely transported over a long distance. In particular, the present invention is a technology for preventing deformation of a specimen by converting a liquid for specimen preservation contained in a vial into a jelly state and thus effectively preventing leakage of the liquid. The present invention has an advantage that the jellification of a liquid for cell fixation filled inside a vial is simple since the liquid for cell fixation is jellified by mixing the selected solution and the selected material.

The present invention relates to methods for diagnosing a disease by determining via multiplex fluorescence in situ hybridization (FISH) whether or not mRNA species and/or at least one miRNA species of disease-associated biomarkers ar present in a sample obtained from a subject, and by determining by multiplex sequencing whether or not said mRNA species of disease-associated biomarkers and/or said miRNA species of disease-associated biomarkers of step (a) are present in said sample. The present invention also relates to kits for performing the methods for diagnosis as described and provided herein as well as use of such kits for performing the methods for diagnosis as described and provided herein.

Provided herein are methods of preserving a biological sample on a first substrate, the methods including: (a) disposing a biological sample on a first substrate; (b) delivering a spacer agent to the first substrate, wherein the spacer agent is applied around the biological sample on the first substrate, thereby generating a chamber around the biological sample; (c) delivering a mounting agent to the chamber; and (d) assembling the first substrate with a second substrate, thereby preserving the biological sample on the first substrate.

A method and system are described for processing tissues according to particular processing protocols that are established based on time-of-flight measurements as a processing fluid is diffused into a tissue sample. In one embodiment, measurement of the time it takes about 70% ethanol to diffuse into a tissue sample is used to predict the time it will take to diffuse other processing fluids into the same or similar tissue samples. Advantageously, the disclosed method and system can reduce overall processing times and help ensure that only samples that require similar processing conditions are batched together.

Certain disclosed embodiments concern a crosslinker having a Formula I

(Polymerizing Group 1).sub.s-(Amino Acid).sub.u-(Polymerizing Group 2).sub.y   Formula I,

where polymerizing group 1 and polymerizing group 2 independently are selected from an acrylate, alkyl acrylate, acrylamide, alkyl acrylamide, acrylonitrile, alkyl acrylonitrile or a (hydroxyalkyl) acrylate; s and y are from 1 to 10, with s and y each typically being 1; the amino acid is any naturally occurring amino acid, any non-naturally occurring amino acid, and any and all combinations thereof; and u is 2 to 100. Crosslinkers may include a naturally occurring amino acid or acids that are selected to provide amino-acid defining functional groups that are zwitterionic at a pH of from 2.5 to 10. Disclosed crosslinkers can also include “internal spacers”, “external spacers,” or both. Crosslinkers according to the present invention are used to make zwitterionic hydrogels that address fouling and bacteria adhesion issues associated with previously known hydrogels. Accordingly, such products are particularly suitable for biomedical applications, such as contact lenses, drug delivery vehicles, tissue engineering platforms, tissue regeneration platforms, catheters, implants and sensors.

Compositions and methods for preparing and using a swellable polymer network for expansion microscopy (ExM) applications. The swellable polymer network can be prepared on demand using external stimuli for ExM applications. A variety of external stimuli can be employed, including light irradiation, pH, temperature, and magnetic fields. Network preparation using light irradiation is particularly advantageous, as it allows tuned control over the network properties through the variation of time and dose of light irradiation. In other words, light irradiation can be employed with a wavelength and intensity that enables a polymer network formation.

The present disclosure provides methods of preparing a biological specimen for imaging analysis, comprising fixing and clearing the biological specimen and subsequently analyzing the cleared biological specimen using microscopy. Also included are methods of quantifying cells, for example, active populations of cells in response to a stimulant. The present disclosure also provides devices for practicing the described methods. A flow-assisted clearing device provides rapid clearing of hydrogel-embedded biological specimens without the need of specialized equipment such as electrophoresis or perfusion devices.

The present disclosure relates to compositions and methods for reversible fixation of biological samples using fixation reagents that form bis-carbamate crosslinks between amine-bearing moieties in biomolecules.

The present invention relates to a method and a kit for rendering a biological material containing melanin suitable for microscopy analysis.