The invention relates to a sterile, disposable, compartmentalized Zavos swim-up column for the swimming-up of spermatozoa, at desired levels of dilution in a media, and a swimming down of spermatozoa into compartments within the column, as well as to a method for the harvesting of those semen samples having particular sperm morphology, motility, progressive motility, speed, sperm concentration, fertilization potential, and a sex ratio. The device comprises a hollow, vertically supported column, having a closed lower end, and an open upper end. A lowermost or first conical member is arranged at the lowermost end of the column, the lowermost conical member having a lowermost periphery in sealing engagement with the bottom of the column. The first conical member has inclined side walls and a truncated uppermost portion defining a peripheral edge which is open within into the column. A second conical member having a lowermost periphery is also attached to the side walls of the column in a sealing manner therebetween. The second conical member also has tapered walls and a truncated uppermost portion defining a peripheral edge which is open to the inside of the column. The area between the inclined walls of each of the conical members and the inner wall of the column define a periconical area which comprises a compartment for collecting sperm prior to harvesting thereof.

BACKGROUND OF THE INVENTION 
1. Field of the Invention 
This invention relates to an apparatus and a method of processing semen and 
allowing its associated spermatozoa to become segregated to permit 
subsequent selection thereof, to increase the number of motile and 
morphological samples to be recovered therefrom. 
2. Prior Art 
A number of methods have been proposed for the processing of semen so as to 
increase its quality and quantity of spermatozoa population of a given 
semen sample. With improved quality and quantity of such spermatozoa, the 
probability of success of Artificial Insemination (AI), In Vitro 
Fertilization (IVF), Intracytoplasmic Sperm Injection (ICSI), and other 
related Assisted Reproductive Clinical Technologies (ARTs), may be 
improved. Some of the methodologies include washing and storage of the 
spermatozoa in media, and the "swim-up" procedure, a technique that takes 
advantage of the swimming abilities of a small percentage of spermatozoa 
within a given sperm population. 
One attempt at the improvement of the collection of spermatozoa is shown in 
U.S. Pat. No. 3,894,529 to Shrimpton, entitled, "Method and Means For 
Controlling the Sex of Mammalian Offspring And Product Therefor". This 
patent discloses a method of separating X and Y sperm so as to control the 
sex of a desired offspring by inseminating an egg with an X or a Y sperm. 
The sperm is separated by applying a buoyant force to a mixture of sperm 
in the nutrient media so that the separation occurs according to the 
density of the sperm. 
U.S. Pat. No. 4,759,344 To Wang, entitled, "Wang's Tubules For Sperm 
Preparations Used For IVF-ET-GIFT and Artificial Inseminations", discloses 
a method and an apparatus for separating sperm having the greatest 
motility. The sperm are allowed to swim upward in a column, where they may 
be collected to provide the strongest samples for insemination. 
Processes such as these are relatively simple, however they are often 
inefficient and maybe susceptible to technician error and/or to laboratory 
variations. 
Further, none of these techniques allow the use of the "swim-up/swim-down" 
principle nor do they permit selectivity based on the various 
characteristics such progressive motility, morphological characteristics, 
X or Y bearing sperm ratio and the like. 
It is an object of the present invention, to provide a method in apparatus 
for the selection of a desired spermatozoa population from a given semen 
sample, to maximize the probability of success when that semen sample is 
utilized for artificial insemination, in vitro fertilization, 
intracytoplasmic sperm injection, and other related assisted reproductive 
clinical technologies. 
It is yet a further object of the present invention, to provide a method 
and an apparatus that will minimize the likelihood of technical error and 
laboratory variations, and permit the selection of a particular population 
of spermatozoa. 
It is yet still a further object of the present invention, to provide a 
method in apparatus to take advantage of the "swim-up/swim-down" principle 
and to permit the entrapment of various classifications of spermatozoa 
based on their various characteristics such as progressive motility, 
morphological characteristics, X or Y bearing sperm ratio and the like. 
BRIEF SUMMARY OF THE INVENTION 
The present invention relates to an apparatus and a method for processing 
semen and allowing its spermatozoa to be selectively sampled, permitting 
an increase in the number of motile and morphologically normal spermatozoa 
in the recovered fractions. 
The apparatus comprises a generally vertically disposed, hollow column 
having a planar or flat lowermost end. The column defines a fluid-tight 
closure having an upper open end. A closure cap is removably securable to 
the open upper end of the column. 
A first or a lowermost truncated conical member having a wide lowermost end 
and a narrower uppermost end is arranged at the bottom of the column. The 
first truncated conical member has a lowermost periphery which sealingly 
engages the lowermost side wall portion of the column, in a sealing 
relationship therewith. The walls of the first truncated conical member 
are arranged at an angle up between about 45 to about 60 degrees with 
respect to the longitude lower axis of the column. The area radially 
outwardly of the longitudinal axis and the tapered walls of the conical 
member is defined as the periconical area. The upper end of the truncated 
conical member is opened to the chamber defined by the interior walls of 
the column. 
A second truncated conical member in a further embodiment, is arranged 
longitudinally above the lowermost conical member. The second conical 
member has a lowermost periphery that engages and is in a sealed 
relationship with the walls of the column. The second truncated conical 
member has an upper periphery through which it is open (it is open at both 
ends), to the chamber defined by the inside walls of the column. The area 
between the walls of the column and the outer side of the second conical 
member is defined as the periconical area or compartment number two. The 
lowermost periphery of the second truncated conical member is preferably 
disposed above the peripheral opening at the truncated portion of the 
first conical member. 
A third and uppermost truncated conical member may be arranged in a mating 
relationship with the inside of the walls of the column, the widest 
portion of the conical member being in a sealed engagement with those 
inside walls of the column at a location spaced above the upper end of the 
second conical member. The third truncated conical member has an uppermost 
opening, which is opened to the inside of the chamber defined by the walls 
of the column. The space between the inside walls of the column and the 
tapered walls of the third truncated conical member is defined as the 
periconical area or compartment number three. 
The area above each respective uppermost periphery or truncated portion of 
each conical member is defined as the epiconical area of its respective 
adjacent conical member. The first conical member is open only at its 
uppermost end. The second conical member and the third conical member are 
each open at both their lower and uppermost ends. Additional truncated 
conical members may be added in a manner similar to the second and third 
conical members, to increase the ability of the apparatus to discriminate 
farther and to improve its accuracy when it is deemed necessary. 
The method of utilization of the present invention involves the placement 
of a semen specimen, (after it has been evaluated for volume, sperm count 
per milliliter, motility, and presence of debris), within the chamber 
defined by the lowermost conical member through its opened upper end. The 
upper surface of the semen sample should be brought to the same level as 
the upper periphery of the open end of the lowermost or first conical 
cylinder. Thereafter, a known volume of isolation media (i.e. "Ham's 
F-10") is drawn to fill the periconical and the epiconical areas of the 
first conical member, if only one such member is utilized within the 
column, or into the epiconical area of the second and third conical 
members, or more, depending on the number of conical members used within 
the inventive device. The isolation media must at least be filled above 
the lip of the upper end of the particular conical member over which the 
sperm is expected to swim to permit the "swim-up/swim-down" process to 
occur. The column then has a closure cap that may be then gently placed 
thereon, and the column and its contents are allowed to sit in a 
controlled environment for about a period of time, (for example, about one 
hour). 
During that period of time, healthy, motile spermatozoa gradually rise or 
swim up from the holding chamber within the lowermost or first conical 
member, through the media, and into the epiconical area of that first 
conical member. Certain spermatozoa will subsequently swim down into the 
periconical area (the first compartment) into the media surrounding the 
lowermost or first conical member. 
Certain spermatozoa specimens will gradually rise and swim-up through the 
upper end of the second conical member and other spermatozoa specimens 
will swim-up through the upper end of the third (or higher) conical 
member. Certain of such specimens will settle in particular periconical 
areas between the walls of the middle or second conical member and the 
column, and the walls of the third (or higher) conical member and the 
inside wall of the column, (if more than one such conical member is used), 
each respective "compartment" or periconical area each defining a 
reservoir for a particular characteristic spermatozoa therewithin. 
At the end of this gradual-rise period, the over-layer of the media in and 
around the top of the semen and within the respective conical members for 
each individual compartment, is removed. The particular spermatozoa 
entrapped in the media within that/those periconical areas (compartments) 
are used for further cases of assisted reproductive technologies. Such 
harvesting or removal of the spermatozoa/media from the compartments 
within the column at the end of the incubation period is done by placing a 
harvesting instrument, such as a needle with a syringe, or a pipette, into 
the respective periconical areas of each conical member, and gently 
aspiring the media until almost all of the overlayed media is removed each 
time. The gradual-rise specimen for each compartment may be used at the 
desired level of dilution in a media of choice, in a centrifuge operation. 
The level of sperm dilution, or the sperm concentration in the resuspended 
preparation, shall be determined by the clinical reproductive purpose for 
that particular spermatozoa. Such gradually-risen spermatozoa may be thus 
utilized for In Vitro Fertilization (IVF), Artificial Insemination (AI), 
Intrauterine Insemination (IUI), Gamete Intra-Fallopian Transfer (GIFT), 
Zygote Intra-Fallopian Transfers (ZIFT), Intra-Cytoplasmic Sperm Injection 
(ICSI), sex selection (shifting the sex ratio via the use of an X or Y 
bearing sperm fraction) and other similar or related clinical assisted 
reproductive procedures or technologies. 
Thus the invention comprises a sterile, disposable, compartmentalized Zavos 
swim-up column for the swimming-up/swimming-down of spermatozoa, at 
desired levels of dilution in a media, for the production of semen samples 
having particular sperm morphology, motility, sperm concentration, 
fertilization potential, and a sex ratio, comprising a hollow, vertically 
supported column, having a closed lower end, and an open upper end. A 
lowermost or first conical member is arranged at the lowermost end of the 
column. The lowermost conical member has a lowermost periphery in sealing 
engagement with the lowermost end of the column. The first conical member 
has inclined side walls and a truncated uppermost portion defining a 
peripheral edge opening within the column. 
A second conical member having a lowermost periphery is attached to side 
walls of said column in a sealing manner therebetween. The second conical 
member has tampered walls and a truncated uppermost portion defining a 
peripheral edge. The second conical member is open to the inside of said 
column. The area between the inclined walls of the conical members, and 
the inner walls of the column define a periconical area that comprises a 
compartment for collecting sperm prior to harvesting thereof. A sealable 
cap is arranged to close the open end of the column. A further conical 
member is arranged within the column, the further conical member defining 
a further periconical compartment or area between the inclined walls of 
the conical member and the walls of the column, for the collection of 
sperm therein, until harvesting thereof. 
The invention includes a method for the collection and harvesting of sperm 
samples of several varying characteristics, may comprise the steps of 
providing a column having a closed lower end and an open upper end, 
placing a first truncated conically shaped member at the lower end of the 
column, placing a second truncated conically shaped member in the column, 
above the first conically shaped member, depositing a sample of sperm 
within the column, within the first conically shaped member, filling the 
column with an isolation media to a level above the upper edge of the 
second truncated conically shaped member, permitting the sperm to swim up 
through the media about the conical member and to swim down into a 
periconical compartment between one of the conical members and the walls 
of the column, and inserting a harvesting device to withdraw media and 
certain sperm from a periconical compartment outwardly of one of the 
conical members. The method may include the steps of inserting a 
harvesting device to withdraw media and certain sperm from a further 
periconical compartment outwardly of a further one of the conical members, 
arranging a third truncated conically shaped member within the column, 
longitudinally above the second conically shaped member; and filling the 
isolation media to a level above the uppermost edge of the third conically 
shaped member, to provide sperm within the first conically shaped member 
yet a further compartment in which to swim up and swim down into. 
Additional conically shaped members may be placed in the column, on top of 
and spaced from the third truncated conical member in a similar 
configuration to the second and third conical members, to increase the 
ability of the apparatus to further discriminate and improve its accuracy 
in selecting higher quality sperm whenever deemed necessary.

DESCRIPTION OF THE PREFERRED EMBODIMENTS 
Referring now to the drawings in detail, and particularly to FIG. 1, there 
is shown the present invention relates to an apparatus and a method for 
processing semen and allowing its spermatozoa to be selectively sampled, 
permitting an increase in the number of motile, progressively motile and 
morphologically normal spermatozoa in the recovered fractions. 
The apparatus comprises a generally vertically disposed, hollow column 10 
having a planar or flat lowermost end 12. The column 10 defines a 
fluid-tight closure having an upper open end 14. A closure cap 16 is 
removably securable to the open upper end 14 of the column 10. 
A first or a lowermost truncated conical member 20 having a wide lowermost 
end 22 and a narrower uppermost end 24 is arranged at the bottom 12 of the 
column 10. The first truncated conical member 22 has a lowermost periphery 
26 which snuggly engages the lowermost inner sidewall portion of the 
column 10, in a sealing relationship therewith. The walls 28 of the first 
truncated conical member 20 are arranged at an angle "A" of between about 
45 to about 60 degrees with respect to the longitudinal axis "L" of the 
column 10. The area radially outwardly of the longitudinal axis and 
outwardly of the tapered walls 28 of the first conical member 20 and 
within the walls of the column 10 is defined as the periconical area "P1". 
The upper end 24 of the first truncated conical member 20 is opened to the 
chamber defined by the interior walls of the column 10, the area 
thereabove being defined as its epiconical area "E1". 
A second truncated conical member 30 in a further embodiment, may be 
arranged longitudinally above the lowermost conical member 20, as shown in 
FIG. 1. The second conical member 30 has a wide lowermost peripheral 
portion 32 that engages and is in a sealed relationship with the walls of 
the column 10. The second truncated conical member 30 has an upper 
peripheral portion 34 through which it is open (open at both ends), to the 
chamber defined by the inside walls of the column 10. The area between the 
walls of the column 10 and the outer side of the second conical member 30 
is defined as the periconical area "P2", as compartment number two. The 
lowermost periphery 32 of the second truncated conical member 30 is 
preferably disposed above the peripheral opening 24 at the truncated 
portion of the first conical member 20, designated "E1". 
A third and uppermost truncated conical member 40 is arranged in a mating 
relationship with the inside of the walls of the column 10, as shown on 
FIG. 1, the lowest and widest periphery 42 of the third conical member 40 
is in a sealed engagement with those inside walls of the column 10 at a 
location spaced above the upper end 34 of the second conical member 30. 
The third truncated conical member 30 has an uppermost opening 44, which 
is opened to the inside of the chamber defined by the walls of the column 
10. The space between the inside walls of the column 10 and the tapered 
walls 46 of the third truncated conical member 40 is defined as the 
periconical area "P3" for compartment number three. 
The area above each respective uppermost periphery or truncated portion of 
each conical member is defined as the epiconical area E1, E2 and E3, for 
its respective adjacent conical member 20, 30 and 40, respectively. The 
first conical member 20 is open only at its uppermost end 24. The second 
conical member 30 and the third conical member 40, and any additional such 
members, are each open at both their lower and uppermost ends. 
The method of utilization of the present invention involves the placement 
of a semen specimen, (after it has been evaluated for volume, sperm count 
per milliliter, motility, and presence of debris), within the chamber 
defined by the lowermost conical member 20 through its opened upper end 
24. The upper surface of the semen sample should be brought to the same 
level as the upper periphery of the open end 24 of the lowermost or first 
conical member 20, as may be seen in FIG. 2. Thereafter, a known volume of 
isolation media (i.e. "Ham's F-10") is drawn to fill the periconical and 
the epiconical areas of the second and third conical members 30 and 40, 
and slightly above, as may be seen in FIG. 3. The column 10 then has a 
closure cap 16, which is then gently placed thereon, and the column 10 and 
its contents are allowed to sit in a controlled environment for a set 
period of time, (for example, about one hour). In any case, the time set 
will be proportional to the number and quality of sperm in the semen to be 
processed. 
During that period of time, healthy, motile spermatozoa gradually rise or 
swim up from the holding chamber within the lowermost or first conical 
member 20, through the media, and into the epiconical area "E1", of that 
first conical member. Certain spermatozoa will subsequently swim down into 
the periconical area (the first compartment P1) into the media surrounding 
the lower-most or first conically shaped member. 
Certain spermatozoa specimens will gradually rise and swim-up through the 
upper end of the second conical member and other spermatozoa specimens 
will swim-up through the upper end of the third conical member. Certain of 
such specimens will settle in particular periconical areas between the 
walls of the middle or second conical member and the column, and the walls 
of the third cone member and the inside wall of the column, each 
respective "compartment" or area each defining a reservoir for a 
particular class of spermatozoa therewithin. 
At the end of this gradual-rise period, the over-layer of the media in and 
around the top of the semen and within the respective conical members for 
each individual compartment, is removed. The particular spermatozoa 
entrapped in the media within those periconical areas (compartments) are 
used for further cases of assisted reproductive technologies. Such 
harvesting or removal of the spermatozoa/media from the column at the end 
of the incubation period is done by placing a harvesting instrument, such 
as a needle with a syringe, or a pipette into the respective periconical 
areas (P3, P2 and P1) of each compartment, and gently aspiring the media 
until almost all of the overlayed media is removed each time. The 
gradual-rise specimen for each compartment may be used at the desired 
level of dilution in a media of choice, in a centrifuge operation. The 
level of sperm dilution, or the sperm concentration in the resuspended 
preparation, shall be determined by the clinical reproductive purpose for 
that particular group of spermatozoa. Such gradually-risen spermatozoa may 
be thus utilized for In Vitro Fertilization (IVF), Artificial Insemination 
(AI), Intrauterine Insemination (IUI), Gamete Intra-Fallopian Transfer 
(GIFT), Zygote Intra-Fallopian Transfers (ZIFT), Intra-Cytoplasmic Sperm 
Injection (ICSI), sex selection (shifting the sex ratio via the use of an 
X or Y bearing sperm fraction) and other assisted reproductive clinical 
procedures.