# US 5,707,860 A · Vehicle for delivery of particles to a sample

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- Number: US 5,707,860 A
- Document ID: US5707860A
- Status: Granted
- Kind: A
- Application number: 08/614,230
- Filed: 1996-03-12
- Granted: 1998-01-13
- Earliest claimed: 1996-03-12
- Assignees: Becton, Dickinson and Company
- Inventors: Matthew P. Collis; Allen S. Reichler
- Web page: https://patentgrep.com/patent/US5707860A

## CPC

- C12Q 1/24: Methods of sampling, or inoculating or spreading a sample; Methods of physically isolating an intact microorganisms
- B01L 3/50825: {Closing or opening means, corks, bungs}
- C12M 47/06: {Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material (lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003)}
- C12M 47/20: {Heating or cooling (heating or cooling apparatus for laboratory uses B01L7/00)}
- C12N 1/04: Preserving or maintaining viable microorganisms (immobilised microorganisms C12N11/00)
- C12Q 1/00: Measuring or testing processes involving enzymes, nucleic acids or microorganisms (measuring or testing apparatus with condition measuring or sensing means, e.g. colony counters, C12M1/34); Compositions therefor; Processes of preparing such compositions
- B01L 2400/0677: phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
- G01N 2035/00267: {Meltable plugs}
- G01N 2333/35: from Mycobacteriaceae (F)
- Y10T 436/25375: Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
- C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- B01L: CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q)
- Y10T: TECHNICAL SUBJECTS COVERED BY FORMER US CLASSIFICATION

## Abstract

The present invention relates to a vehicle for delivery of particles to a sample of cells. The vehicle includes a barrier to retain the particles, which barrier in one embodiment is a frangible, dissolvable or meltable material. Once released into the sample, the particles are useful in methods to lyse or disrupt cells or in methods to separate cellular components from one another if the cells in the sample are already lysed or disrupted.

## Claims

### Claim 1

A vehicle for delivery of particles to a sample containing cells comprising a barrier which retains the particles until said particles are released into the sample, said particles being sufficiently non-dissolvable to remain undissolved in the sample for a time sufficient to disrupt the cells and render accessible nucleic acids therefrom when the sample is agitated or sonicated.

### Claim 2 (depends on 1)

The vehicle of claim 1 wherein the barrier comprises a frangible material which releases the particles after breaking.

### Claim 3 (depends on 2)

The vehicle of claim 2 wherein at least one particle is of sufficient size to cause the breaking of the barrier and the remainder of the particles are of a lesser size.

### Claim 4 (depends on 1)

The vehicle of claim 1 wherein the particles are glass beads.

### Claim 5 (depends on 1)

The vehicle of claim 1 wherein the particles are zirconium/silicate beads.

### Claim 6 (depends on 3)

The vehicle of claim 3 wherein said at least one particle is a glass bead and the remainder of the particles are zirconium/silicate beads.

### Claim 7 (depends on 1)

The vehicle of claim 1 wherein the barrier comprises a material which dissolves when contacted with the sample.

### Claim 8 (depends on 1)

The vehicle of claim 1 wherein the barrier comprises a material which melts at a temperature greater than about 80.degree. C.

### Claim 9 (depends on 1)

A method for rendering cellular components accessible comprising the steps of:
(a) adding to a sample containing cells, the vehicle of claim 1;
(b) disrupting the cells to cause release of cellular components;
(c) causing the particles to be released from the vehicle; and
(d) agitating the sample sufficiently such that nucleic acids are separated from other cellular components.

### Claim 10 (depends on 8)

The method of claim 8 wherein the cells are disrupted by heating the sample at a temperature and for a time sufficient to render infectious organisms in the sample noninfectious.

### Claim 11 (depends on 8)

The method of claim 8 wherein the particles are caused to be released from the vehicle by impact of a particle with the barrier.