We claim:
1. A method for detecting lymphocyte activation comprising the steps of:
incubating a sample containing a mixed population of cell types including a plurality of subsets of lymphocytes, wherein each subset includes lymphocytes with characteristic determinants that distinguish one subset from another, with at least one inducing agent selected from the group consisting of mitogens, antigens and co-stimulants;
separating a selected subset of lymphocytes from said sample;
lysing lymphocytes in said selected subset to release an intracellular component selected from the group consisting of ATP, NADP, and PCNA;
detecting a level of said intracellular component; and,
assessing lymphocyte activation for said selected subset of lymphocytes from said level of intracellular component detected in said detecting step, wherein the total time required for performing all steps is no greater than one hour.
2. The method of claim 1 wherein said at least one inducing agent comprises a co-stimulant.
3. A method for detecting lymphocyte activation comprising the steps of:
incubating a sample containing a mixed population of cell types including a plurality of subsets of lymphocytes, wherein each subset includes lymphocytes with characteristic determinants that distinguish one subset from another, with at least one inducing agent selected from the group consisting of mitogens, antigens and co-stimulants;
separating a selected subset of lymphocytes from said sample;
lysing lymphocytes in said selected subset to release an intracellular component selected from the group consisting of ATP, NADP, and PCNA;
detecting a level of said intracellular component; and,
assessing lymphocyte activation for said selected subset of lymphocytes from said level of intracellular component detected in said detecting step, wherein the total time required for performing all steps is less than three hours.
4. The method of claim 3 wherein said at least one inducing agent comprises a co-stimulant.
5. A method for detecting lymphocyte activation comprising the steps of:
incubating a sample containing a mixed population of cell types including a plurality of subsets of lymphocytes, wherein each subset includes lymphocytes with characteristic determinants that distinguish one subset from another, with at least one inducing agent selected from the group consisting of mitogens, antigens and co-stimulants;
separating a selected subset of lymphocytes from said sample;
lysing lymphocytes in said selected subset to release an intracellular component selected from the group consisting of ATP, NADP, and PCNA;
detecting a level of said intracellular component; and,
assessing lymphocyte activation for said selected subset of lymphocytes from said level of intracellular component detected in said detecting step, wherein the total time required for performing all steps is less than four hours.
6. The method of claim 5 wherein said at least one inducing agent comprises a co-stimulant.
7. A method for detecting lymphocyte activation comprising the steps of:
incubating a sample containing a mixed population of cell types including a plurality of subsets of lymphocytes, wherein each subset includes lymphocytes with characteristic determinants that distinguish one subset from another, with at least one inducing agent selected from the group consisting of mitogens, antigens and co-stimulants;
separating a selected subset of lymphocytes from said sample;
lysing lymphocytes in said selected subset to release an intracellular component selected from the group consisting of ATP, NADP, and PCNA;
detecting a level of said intracellular component; and,
assessing lymphocyte activation for said selected subset of lymphocytes from said level of intracellular component detected in said detecting step, wherein the total time required for performing all steps is less than 24 hours.
8. The method of claim 7 wherein said at least one inducing agent comprises a co-stimulant.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1.-68. (canceled)
69. A fluid pressure pulse generating apparatus for downhole drilling comprising:
(a) a fluid pressure pulse generator;
(b) a pulser assembly longitudinally adjacent the fluid pressure pulse generator with a fluid flow channel extending between adjacent surfaces thereof, the pulser assembly comprising a pulser assembly housing that houses a motor and a driveshaft extending from the motor out of the pulser assembly housing and coupling with the fluid pressure pulse generator; and
(c) a primary seal surrounding a portion of the driveshaft and configured to prevent drilling fluid from entering the pulser assembly housing and lubrication liquid from leaving the pulser assembly housing when the fluid pressure pulse generating apparatus is positioned downhole,
wherein the fluid flow channel defines at least a portion of a flow path for the drilling fluid which flows from external the pulser assembly to the primary seal when the fluid pressure pulse generating apparatus is positioned downhole, whereby the adjacent surfaces of the pulser assembly and the fluid pressure pulse generator are configured such that the fluid flow channel comprises a tortuous flow path.
70. The apparatus of claim 69, wherein the fluid flow channel includes a plurality of changes in direction.
71. The apparatus of claim 69, wherein the fluid flow channel comprises a restricted section and an expanded section, whereby the cross sectional area of the restricted section is less than the cross sectional area of the expanded section.
72. The apparatus of claim 71, wherein the expanded section comprises an expansion chamber having an increased volume compared to the volume of the restricted section.
73. The apparatus of claim 69, wherein the pulser assembly further comprises a journal bearing surrounding a portion of the driveshaft with a gap between an internal surface of the journal bearing and an external surface of the driveshaft, the journal bearing being positioned on the driveshaft between the coupling with the pressure pulse generator and the primary seal, wherein the gap defines at least a portion of the flow path for the drilling fluid.
74. The apparatus of claim 73, wherein the volume of drilling fluid flowing through the gap is restricted compared to the volume of drilling fluid in the flow path before andor after the gap.
75. The apparatus of claim 73, further comprising a primary sealing cylinder fitted on a portion of the driveshaft such that the primary seal seals against an outer sealing surface of the primary sealing cylinder and the journal bearing aligns with the outer sealing surface such that the gap is between the outer sealing surface and the external surface of the journal bearing.
76. The apparatus of claim 75, wherein the primary sealing cylinder is configured to releasably fit on the driveshaft.
77. The apparatus of claim 76, wherein the driveshaft comprises a first annular shoulder and the primary sealing cylinder is positioned between the first annular shoulder and the fluid pressure pulse generator to releasably secure the primary sealing cylinder on the driveshaft.
78. The apparatus of claim 73, wherein the flow path for the drilling fluid further comprises a fluid expansion chamber positioned between the journal bearing and the primary seal, wherein the volume of drilling fluid in the fluid expansion chamber is greater than the volume of drilling fluid in the gap.
79. The apparatus of claim 73, wherein the pulser assembly further comprises a journal bearing housing enclosing the journal bearing, the journal bearing housing configured to releasably mate with the pulser assembly housing.
80. The apparatus of claim 79, wherein the journal bearing housing comprises a cylindrical section which surrounds a circular section of the fluid pressure pulse generator, the circular section of the fluid pressure pulse generator configured to rotate within the cylindrical section of the journal bearing housing, whereby the fluid flow channel extends between an internal surface of the cylindrical section and an external surface of the circular section.
81. The apparatus of claim 69, wherein the pulser assembly housing comprises a cylindrical section which surrounds a circular section of the fluid pressure pulse generator, the circular section of the fluid pressure pulse generator configured to rotate within the cylindrical section of the pulser assembly housing, whereby the fluid flow channel extends between an internal surface of the cylindrical section and an external surface of the circular section.
82. The apparatus of claim 69, further comprising a pressure compensation device comprising:
(a) a membrane support having a longitudinally extending bore therethrough for receiving a driveshaft of the fluid pressure pulse generating apparatus; and
(b) a longitudinally extending membrane system comprising a longitudinally extending outer membrane sleeve and a longitudinally extending inner membrane sleeve with the inner membrane sleeve positioned inside the outer membrane sleeve,
wherein the membrane system is sealed to the membrane support to allow flexing of the membrane system in response to fluid pressure on either an inner longitudinal surface of the membrane system or an outer longitudinal surface of the membrane system and to prevent fluid on the inner longitudinal surface mixing with fluid on the outer longitudinal surface,
said device surrounding a portion of the driveshaft and positioned in the pulser assembly housing so that the outer longitudinal surface of the membrane system is exposed to the drilling fluid flowing external to the pulser assembly housing when the fluid pressure pulse generating apparatus is positioned downhole and the inner longitudinal surface of the membrane system is exposed to the lubrication liquid contained inside the pulser assembly housing.
83. The apparatus of claim 82, wherein the pulser assembly housing comprises a plurality of apertures extending therethrough, the plurality of apertures being in fluid communication with the outer longitudinal surface of the membrane system.
84. The apparatus of claim 83, further comprising a longitudinally extending drilling fluid chamber adjacent the outer longitudinal surface of the membrane system, the drilling fluid chamber being in fluid communication with the plurality of apertures.
85. The apparatus of claim 69, further comprising a back up seal enclosed by the pulser assembly housing and surrounding a portion of the driveshaft between the primary seal and the motor, the back up seal configured to prevent the lubrication liquid on a primary seal side of the back up seal from mixing with the lubrication liquid on a motor side of the back up seal.
86. The apparatus of claim 85, further comprising a back up sealing cylinder fitted on a portion of the driveshaft such that the back up seal seals against an outer sealing surface of the back up sealing cylinder.
87. The apparatus of claim 86, wherein the back up sealing cylinder is configured to releasably fit on the driveshaft.
88. The apparatus of claim 87, further comprising a back up seal housing enclosing the back up seal and the back up seal cylinder, the back up seal housing comprising a first section and a second section configured to releasably mate with the first section, wherein the driveshaft comprises a second annular shoulder and the back up sealing cylinder is positioned between the second annular shoulder and an internal surface of the back up seal housing to releasably secure the back up sealing cylinder on the driveshaft.
89. The apparatus of claim 88, further comprising a retention nut surrounding a portion of the driveshaft and configured to releasably secure the first section and the second section of the back up seal housing together so as to releasably secure the back up sealing cylinder on the driveshaft.
90. The apparatus of claim 85, further comprising a back up seal housing enclosing the back up seal.
91. The apparatus of claim 90, wherein the back up seal housing comprises a first section and a second section configured to releasably mate with the first section.
92. The apparatus of claim 88, further comprising a thrust bearing surrounding a portion of the driveshaft and enclosed by the back up seal housing.
93. The apparatus of claim 88, further comprising a first thrust bearing surrounding a portion of the driveshaft on one side of the back up seal and a second thrust bearing surrounding a portion of the driveshaft on an opposed side of the back up seal, the first and second thrust bearings being enclosed by the back up seal housing.
94. The apparatus of claim 85, further comprising a spring positioned longitudinally adjacent and in communication with the back up seal for spring loading the back up seal.
95. The apparatus of claim 85, wherein the lubrication liquid on the primary seal side of the back up seal has a different composition to the lubrication liquid on the motor side of the back up seal.
96. The apparatus of claim 95, wherein the lubrication liquid on the primary seal side of the back up seal has a higher viscosity than the lubrication liquid on the motor side of the back up seal.
97. The apparatus of claim 95, wherein the lubrication liquid on the primary seal side of the back up seal has a lower thermal expansion than the lubrication liquid on the motor side of the back up seal.
98. The apparatus of claim 69, wherein the primary seal is positioned uphole of the entrance to the fluid flow channel.