Abstrict A seed crusher e.g. useful in methods for high throughput analysis
of seeds, comprises a horizontal well plate with at least one well
for receiving a tube; an open-topped tube for lining each well wherein
each tube is oriented for receiving a vertical reciprocating pin;
a horizontal die plate with a vertical pin located to reciprocate
in each tube; a removable separator for use in each tube to separate
a reciprocating pin from a seed at the bottom of said tube and a
press for bringing together the die plate and well plate to allow
a pin to enter each tube.
Claims 1. A seed crusher, comprising (a) a horizontal well plate with
at least one well for receiving a tube; (b) an open-topped tube
for lining each well wherein each tube is oriented for receiving
a vertical reciprocating pin; (c) a horizontal die plate with a
vertical pin located to reciprocate in each tube; (d) a removable
separator for use in each tube to separate a reciprocating pin from
a seed at the bottom of said tube; and (e) a press for bringing
together the die plate and well plate to allow a pin to enter each
tube.
2. The seed crusher of claim 1 wherein said well plate is moveable
and said die plate is fixed.
3. The seed crusher of claim 1 wherein said well plate is fixed
and said die plate is moveable.
4. The seed crusher of claim 1 wherein said well plate has a plurality
of wells.
5. The seed crusher of claim 4 comprising a plurality of tubes
fixed in an array.
6. A method of seed extraction using the seed crusher of claim
5 comprising the steps of: (a) placing at least one seed and a removable
separator in each tube; wherein said separator transfers the pressure
from said pin to the seed at the bottom of the tube; (b) operating
the seed crusher to reciprocate the pin in said tube to crush the
seed in the tube; (c) delivering extraction medium in each tube
wherein said medium can dissolve seed components; and (d) recovering
the extraction medium from each tube.
7. The method of claim 6 wherein said tube is shaken after delivering
the extraction medium in the tube and before recovering the extraction
medium from the tube.
8. The method of claim 7 where in said extraction medium is delivered
in a separate tube after recovering the extraction medium from the
tube.
Description [0001] This application claims the benefit of U.S. application
No. 60/549435 filed Mar. 02 2004.
FIELD OF THE INVENTION
[0002] The present invention describes a seed crusher and its use
in extraction of seed components.
BACKGROUND OF THE INVENTION
[0003] One of the goals of plant genetic engineering or conventional
plant breeding is to produce plants with agronomically, horticulturally
or economically important characteristics or traits. The testing
of plant tissue for producing plants with improved desired characteristics
often requires extraction of metabolites and other bio-molecules
from soft tissue such as leaf or hard tissue such as seed. The first
step in extraction from seed involves crushing which requires significant
mechanical force for hard seeds. The manual application of force
required for seed crushing in a high throughput screening is tedious,
cumbersome, and usually a source of repetitive stress injuries.
SUMMARY OF THE INVENTION
[0004] This invention provides a seed crusher adapted for crushing
seeds in tubes which can serve as containers for further analytical
processing. More particularly, a seed crusher comprises; (a) a horizontal
well plate with at least one well for receiving a tube; (b) an open-topped
tube for lining each well, where each tube is oriented for receiving
a vertical reciprocating pin; (c) a horizontal die plate with a
vertical pin located to reciprocate in each tube; (d) a removable
separator for use in each tube to separate a reciprocating pin from
a seed at the bottom of said tube; and (e) a press for bringing
together the die plate and well plate to allow a pin to enter each
tube. In one embodiment the well plate is moveable and the die plate
is fixed. In another embodiment the well plate is fixed and the
die plate is moveable. In another embodiment the well plate has
a plurality of wells and the seed crusher comprises a plurality
of wells. Preferably the wells and tubes are in an array, which
allows the plurality of tubes to be in a fixed structure. When the
seed crusher comprises a plurality of wells and tubes, the die plate
has a plurality of reciprocating pins, i.e. a pin aligned for reciprocating
in each tube. Each well serves to hold and support the bottom of
a tube, e.g. to prevent breakage from pushing a crushed seed through
the bottom of a tube. In one embodiment the most of the tube can
be located in a deep well. In another embodiment only the bottom
of the tube lines the well.
[0005] The invention also provides a method of crushing seed by
placing one or more seeds along with one or more removable separators
in each tube and operating the seed crusher to reciprocate the pin
in the tube. The invention further proves a method of extracting
one or more constituents from crushed seed by the addition of an
extraction medium in a tube with crushed seed and recovering the
extraction medium from the tube.
BRIEF DESCRIPTION OF THE DRAWINGS
[0006] FIG. 1 is an illustration of components of a seed crusher.
[0007] FIG. 2 is an illustration of a method of using a seed crusher.
DETAILED DESCRIPTION
[0008] With reference to FIGS. 1 and 2 there is shown a horizontal
plate 1 having a well 2 supporting a tube 3. After a seed 6 and
removable separator 7 is placed in a tube 3 a pin 5 attached to
a die plate 4 reciprocates in the tube 3 sufficiently to crush the
seed 6.
[0009] The function of the horizontal plate is to provide a strong
base for tube-supporting wells. The horizontal plant can be static
or moveable. The horizontal plate can be made of any material, which
can withstand the pressure needed to crush seed in the tube, e.g.
about 20-40 megaPascals [3000 to 6000 pounds per square inch].
A well plate can effectively be made from metal, e.g. steel or aluminum,
or plastic, e.g. reinforced plastic. Each well in a well plate is
designed to provide a relatively snug fit to conform to the base
of a tube in order to prevent the brakeage of the tube. As tubes
are preferably removable the well should be designed to facilitate
easy removal of a tube.
[0010] An open-topped tube for use in the seed crusher has one
open end for inserting and removing materials, e.g. seeds, separators
and reciprocating pins, and one closed end to contain the contents
of the tube. The shape of the bottom of the tube is such that it
matches the shape of the well on the plate to provide a support
to bottom of the tube by the well in the horizontal plate. The material
of the tube is chosen to sustain the force applied by the reciprocating
pin in order to maintain the integrity of the tube. Plastic such
as polypropylene is a useful material of construction for tubes.
In one embodiment a single tube fits a single well in the horizontal
plate. In other embodiment an array of a plurality of tubes is in
a unit, e.g. as a "multi-well" plate or a box with multiple
tubes to fit corresponding multiple wells in the horizontal plate.
[0011] The die plate with a pin for each tube is designed to transfer
the pressure from the die plate to pins contacting separators contacting
seeds in the bottom of a tube. Each pin of the die plate is designed
to reciprocate in a tube. The end of the pin that comes in contact
with the separator is designed to transfer desired pressure to one
or more seeds via one or more separators in the tube.
[0012] To effect seed crushing the horizontal plate and die plate
are aligned so that, when the plates are pressed together, a pin
will enter each tube as shown in FIG. 2 allowing a pin to press
a separator onto a seed at the bottom of a tube. The plates can
be mounted in a press where a compressing force is applied by hydraulic
pistons, air pistons, electric driven screws, mechanical lever or
mechanical driven screws. For rapid seed crushing a press with fast
acting piston is preferred.
[0013] The amount of the force needed to crush one or more seeds
is variable and depends on the variety of seed, size of the seed,
shape of the seed, moisture content of the seed and the interaction
between the pin and the seed via the separator. For the present
invention a desired press will have properties where the amount
of force can be varied to that required for adequately crushing
the type of seed being processed. In one embodiment, the press is
configured to apply a desired force to one or more seeds via a pin
and one or more separators where the die plate is moveable and the
horizontal plate is fixed. In other embodiment, the press is configured
to apply a desired force to one or more seeds via a pin and one
or more separators where the die plate is fixed and the horizontal
plate is moveable.
[0014] The present invention also describes a method using the
seed crusher for extraction of seed content. Extraction of components
from seeds is done to perform further analysis of seed composition.
Modem methods of analysis are very sensitive and are negatively
impacted by cross contaminations. One or more separators are placed
in the tube to avoid the contamination of the reciprocating pin
and also for proper distribution of pressure on one or more seeds
placed in the tube. The separator can be of any shape as long as
it meets above criteria. A useful shape of a separator is spherical.
One or more separators can be are placed between one or more seeds
and the pin.
[0015] After crushing the seed by operation of the seed crusher,
the pins are reciprocated out of the tubes and an extraction medium
is delivered into each tube with crushed seed followed by the recovery
of the extraction medium. Depending on the analysis the extraction
medium can be a solvent, which is capable of dissolving the seed
component under analysis; for example, an organic solvent will be
a desired solvent if seed oil is the component under analysis. If
seed component is a water-soluble salt, then the extraction medium
can be water-based. The extraction medium can be delivered in the
tube by any means capable of handling liquid such as a pipette,
a dropper or a liquid handling robot. For increasing the efficiency
of seed extraction, the extraction medium can be agitated along
with the crushed seed content in the tube before the recovery of
extraction medium from the tube. |