339:, due to their ability to maintain constant pressure throughout the piston stroke. As the French Press, which is operated by hydraulic pressure, is capable of over 90% lysis of most commonly used cell types it is often taken as the gold standard in lysis performance and modern machines are often compared against it not only in terms of lysis efficiency but also in terms of safety and ease of use. Some manufacturers are also trying to improve on the traditional design by altering properties within these machines other than the pressure driving the sample through the orifice. One such example is Constant Systems, who have recently shown that their Cell Disruptors not only match the performance of a traditional French Press, but also that they are striving towards attaining the same results at a much lower power.
343:
rupture (lysis) of cells and tissues from human, animal, plant, and microbial sources, and the inactivation of pathogens. PCT-enhanced systems (instruments and consumables) address some challenging problems inherent in biological sample preparation. PCT advantages include: (a) extraction and recovery of more membrane proteins, (b) enhanced protein digestion, (c) differential lysis in a mixed sample base, (d) pathogen inactivation, (e) increased DNA detection, and (f) exquisite sample preparation process control.
119:
331:, or French Press for short. This method was developed by Charles Stacy French and utilises high pressure to force cells through a narrow orifice, causing the cells to lyse due to the shear forces experienced across the pressure differential. While French Presses have become a staple item in many microbiology laboratories, their production has been largely discontinued, leading to a resurgence in alternate applications of similar technology.
22:
315:
biological samples containing a significant fraction of water become brittle at extremely cold temperatures. This technique was first described by
Smucker and Pfister in 1975, who referred to the technique as cryo-impacting. The authors demonstrated cells are effectively broken by this method, confirming by phase and electron microscopy that breakage planes cross cell walls and cytoplasmic membranes.
319:
require less manual effort, give good sample recovery and are easy to clean between samples. Advantages of this technique are higher yields of proteins and nucleic acids from small, hard tissue samples - especially when used as a preliminary step to mechanical or chemical/solvent cell disruption methods mentioned above.
342:
Pressure
Cycling Technology ("PCT"). PCT is a patented, enabling technology platform that uses alternating cycles of hydrostatic pressure between ambient and ultra-high levels (up to 90,000 psi) to safely, conveniently and reproducibly control the actions of molecules in biological samples, e.g., the
334:
Modern physical cell disruptors typically operate via either pneumatic or hydraulic pressure. Although pneumatic machines are typically lower cost, their performance can be unreliable due to variations in the processing pressure throughout the stroke of the air pump. It is generally considered that
350:
cell suspension, such as particle size, viscosity, protein yield and enzyme activity. In recent years the
Microfluidizer method has gained popularity in cell disruption due to its ease of use and efficiency at disrupting many different kinds of cells. The Microfluidizer technology was licensed from
266:
All high energy bead beating machines warm the sample about 10 degrees per minute. This is due to frictional collisions of the beads during homogenization. Cooling of the sample during or after bead beating may be necessary to prevent damage to heat-sensitive proteins such as enzymes. Sample warming
318:
The technique can be done by using a mortar and pestle cooled to liquid nitrogen temperatures, but use of this classic apparatus is laborious and sample loss is often a concern. Specialised stainless steel pulverizers generically known as Tissue
Pulverizers are also available for this purpose. They
239:
Successful bead beating is dependent not only on design features of the shaking machine (which take into consideration shaking oscillations frequency, shaking throw or distance, shaking orientation and vial orientation), but also the selection of correct bead size (0.1–6 mm (0.004–0.2 in)
314:
Samples with a tough extracellular matrix, such as animal connective tissue, some tumor biopsy samples, venous tissue, cartilage, seeds, etc., are reduced to a fine powder by impact pulverization at liquid nitrogen temperatures. This technique, known as cryopulverization, is based on the fact that
461:
the sample or produce unwanted damage. There is no need to watch for a peak between enzyme activity and percent disruption. Since nitrogen bubbles are generated within each cell, the same disruptive force is applied uniformly throughout the sample, thus ensuring unusual uniformity in the product.
382:
Viscosity changes are also often observed when disrupting cells. If the cell suspension viscosity is high, it can make downstream handling—such as filtration and accurate pipetting—quite difficult. The viscosity changes observed with a
Microfluidizer are relatively low, and decreases with further
386:
In contrast to other mechanical disruption methods the
Microfluidizer breaks the cell membranes efficiently but gently, resulting in relatively large cell wall fragments (450 nm), and thus making it easier to separate the cell contents. This can lead to shorter filtration times and better
373:
Many proteins are extremely temperature-sensitive, and in many cases can start to denature at temperatures of only 4 degrees
Celsius. Within the microchannels, temperatures exceed 4 degrees Celsius, but the machine is designed to cool quickly so that the time the cells are exposed to elevated
221:, and can yield breakage of well over 50% (up to 95%). It has the advantage over other mechanical cell disruption methods of being able to disrupt very small sample sizes, process many samples at a time with no cross-contamination concerns, and does not release potentially harmful
289:
rotor inside a 15, 50 or 200 ml chamber to agitate the beads. In this configuration, the chamber can be surrounded by a static cooling jacket. Using this same rotor/chamber configuration, large commercial machines are available to process many liters of
445:
is preferred because of its non-reactive nature and because it does not alter the pH of the suspending medium. In addition, nitrogen is preferred because it is generally available at low cost and at pressures suitable for this procedure.
153:
methods allows the study and manufacture of relevant molecules. Except for excreted molecules, cells producing molecules of interest must be disrupted. This page discusses various methods. Another method of disruption is called
639:
The photochemical reduction process in photosynthesis. In
Symposia of the Society for Experimental Biology. C. S. French, H. W. Milner. V. Carbon Dioxide Fixation and Photosynthesis. 232-250. Cambridge University Press, New
417:
and mechanical homogenizing methods and compares favorably to the controlled disruptive action obtained in a PTFE and glass mortar and pestle homogenizer. While other disruptive methods depend upon friction or a mechanical
186:. First developed by Tim Hopkins in the late 1970s, the sample and bead mix is subjected to high level agitation by stirring or shaking. Beads collide with the cellular sample, cracking open the cell to release the
703:
agerkvist, Irene, and Sven-Olof Enfors.”Characterization Of E. Coli Cell
Disintegrates from a Bead Bill and High Pressure Homogenizer.”Biotechnology and bioengineering Biotechnol.Bioeng.36.11 (1990):1083-089.Web.
694:
agerkvist, Irene, and Sven-Olof Enfors.”Characterization Of E. Coli Cell
Disintegrates from a Bead Bill and High Pressure Homogenizer.”Biotechnology and bioengineering Biotechnol.Bioeng.36.11 (1990):1083-089.Web.
402:. Then, when the gas pressure is suddenly released, the nitrogen comes out of the solution as expanding bubbles that stretch the membranes of each cell until they rupture and release the contents of the cell.
378:
25 ms-40 ms). Because of this effective temperature control, the Microfluidizer yields higher levels of active proteins and enzymes than other mechanical methods when the proteins are temperature-sensitive.
228:
In the simplest example of the method, an equal volume of beads are added to a cell or tissue suspension in a test tube and the sample is vigorously mixed on a common laboratory
263:
also shorten process times, as do Bead Dispensers designed to quickly load beads into multiple vials or microplates. Pre-loaded vials and microplates are also available.
259:. Differing from conventional machines, it agitates the beads using a vortex motion at 20,000 oscillations per minute. Larger bead beater machines that hold deep-well
630:
The photochemical activity of isolated chloroplasts. C. S. French, H. W. Milner, M. L. Koenig, and F. D. H. Macdowall. Carn. Inst. Wash. Yearb. 1948; 47:91-93
621:
Liquid Nitrogen Cryo-Impacting: a New Concept for Cell Disruption. Richard A. Smucker, Robert M. Pfister. Appl Microbiol. 1975 September; 30(3): 445–449.
255:. Cell disruption is complete in 1–3 minutes of shaking. Significantly faster rates of cell disruption are achieved with a bead beater variation called
422:
action that generate heat, the nitrogen decompression procedure is accompanied by an adiabatic expansion that cools the sample instead of heating it.
746:
558:
251:. The sample and tiny beads are agitated at about 2000 oscillations per minute in specially designed reciprocating shakers driven by high power
661:"Hands-free Sample Homogenization and Protein Extraction from Small Tissue Biopsy Samples using Pressure Cycling Technology and PCT µPestle"
722:
712:
Evaluation of the Microfluidizer for Cell Disruption of Yeast and Chlorella by E. Uera-Santos, C.D. Copple, EA Davis and WG. Hagar.
685:
Evaluation of the Microfluidizer for Cell Disruption of Yeast and Chlorella by E. Uera-Santos, C.D. Copple, EA Davis and WG. Hagar.
398:
For nitrogen decompression, large quantities of nitrogen are first dissolved in the cell under high pressure within a suitable
86:
58:
583:
202:
or subcellular preparations. The method, often called "bead beating", works well for all types of cellular material - from
481:, and produce uniform and repeatable homogenates without subjecting the sample to extreme chemical or physical stress.
454:
65:
351:
a company called Arthur D. Little and was first developed and utilized in the 1980s, initially starting as a tool for
335:
hydraulic machines offer superior lysing ability, especially when processing harder to break samples such as yeast or
105:
39:
425:
The blanket of inert nitrogen gas that saturates the cell suspension and the homogenate offers protection against
370:
are generated that rupture the cells. This method of cell lysis can yield breakage of over 90% of E. coli cells.
375:
72:
660:
363:
43:
346:
The Microfluidizer method used for cell disruption strongly influences the physicochemical properties of the
54:
458:
247:
In most laboratories, bead beating is done in batch sizes of one to twenty-four sealed, plastic vials or
532:
328:
195:
466:
659:
Shao, Shiying; Gross, Vera; Yan, Wen; Guo, Tiannan; Lazarev, Alexander; Aebersold, Ruedi (2015).
32:
327:
Since the 1940s high pressure has been used as a method of cell disruption, most notably by the
522:
336:
179:
776:
79:
295:
285:
A different bead beater configuration, suitable for larger sample volumes, uses a rotating
8:
233:
130:
419:
187:
723:"Parr Instruments - Cell Disruption Vessel, 1 gallon internal volume - John Morris"
260:
232:. While processing times are slow, taking 3–10 times longer than that in specialty
183:
438:
399:
291:
279:
248:
608:
587:
488:
and other membrane-bound cells. It has also been used successfully for treating
649:
Under Pressure, M. Lougher, European Biopharmaceutical Review, July 2016; 12-16
430:
355:
creation. It has since been used in other applications such as cell disruption
252:
155:
134:
508:
and other materials with tough cell walls do not respond well to this method.
770:
474:
434:
390:
Microfluidizer technology scales from one milliliter to thousands of liters.
207:
199:
556:
286:
267:
can be controlled by bead beating for short time intervals with cooling on
229:
211:
191:
178:
beads, 0.1–2 mm (0.004–0.08 in) in diameter, mixed with a sample
150:
537:
478:
450:
240:
diameter), bead composition (glass, ceramic, steel) and bead load in the
527:
489:
470:
414:
367:
118:
493:
485:
426:
410:
275:
166:
A common laboratory-scale mechanical method for cell disruption uses
492:, for releasing virus from fertilized eggs and for treating fragile
21:
442:
356:
352:
303:
272:
222:
171:
146:
505:
501:
406:
236:, it works well for easily disrupted cells and is inexpensive.
517:
497:
347:
299:
218:
215:
203:
175:
167:
241:
145:
The production of biologically interesting molecules using
271:
between each interval, by processing vials in pre-chilled
268:
496:. It is not recommended for untreated bacterial cells.
294:. Currently, these machines are limited to processing
441:
and compressed air have been used in this technique,
484:
The method is particularly well suited for treating
366:with fixed geometry, and an intensifier pump, high
359:, and solid particle size reduction, among others.
46:. Unsourced material may be challenged and removed.
557:EMBL - Office of Information and Public Affairs.
768:
322:
658:
214:tissues. It is the most widely used method of
405:Nitrogen decompression is more protective of
429:of cell components. Although other gases:
393:
282:through the machine during bead beating.
106:Learn how and when to remove this message
667:. Institute of Molecular Systems Biology
453:substances are not exposed to continued
117:
462:Cell-free homogenates can be produced.
383:additional passes through the machine.
769:
129:is a method or process for releasing
309:
44:adding citations to reliable sources
15:
13:
469:cells and tissues, release intact
14:
788:
609:"BioSpec Products • Bead Loaders"
374:temperatures is extremely short (
20:
739:
715:
706:
697:
688:
31:needs additional citations for
679:
652:
643:
633:
624:
615:
601:
576:
550:
161:
1:
543:
323:High Pressure Cell Disruption
190:. Unlike some other methods,
7:
511:
387:centrifugation separation.
10:
793:
533:Homogenization (chemistry)
329:French Pressure Cell Press
140:
465:The technique is used to
122:Laboratory cell disruptor
665:Pressure Biosciences Inc
188:intracellular components
751:Parr Instrument Company
727:www.johnmorrisgroup.com
198:resulting in excellent
559:"Protein Purification"
523:Ultrasonic homogenizer
394:Nitrogen decompression
337:Gram-positive bacteria
123:
296:unicellular organisms
121:
131:biological molecules
40:improve this article
194:is moderate during
278:or by circulating
124:
584:"Bead Dispensers"
477:, release labile
310:Cryopulverization
261:microtiter plates
116:
115:
108:
90:
55:"Cell disruption"
784:
761:
760:
758:
757:
743:
737:
736:
734:
733:
719:
713:
710:
704:
701:
695:
692:
686:
683:
677:
676:
674:
672:
656:
650:
647:
641:
637:
631:
628:
622:
619:
613:
612:
605:
599:
598:
596:
595:
586:. Archived from
580:
574:
573:
571:
569:
554:
249:centrifuge tubes
234:shaking machines
225:in the process.
192:mechanical shear
184:aqueous solution
111:
104:
100:
97:
91:
89:
48:
24:
16:
792:
791:
787:
786:
785:
783:
782:
781:
767:
766:
765:
764:
755:
753:
745:
744:
740:
731:
729:
721:
720:
716:
711:
707:
702:
698:
693:
689:
684:
680:
670:
668:
657:
653:
648:
644:
638:
634:
629:
625:
620:
616:
607:
606:
602:
593:
591:
582:
581:
577:
567:
565:
555:
551:
546:
514:
449:Once released,
439:carbon monoxide
400:pressure vessel
396:
325:
312:
298:such as yeast,
292:cell suspension
280:gaseous coolant
253:electric motors
164:
143:
127:Cell disruption
112:
101:
95:
92:
49:
47:
37:
25:
12:
11:
5:
790:
780:
779:
763:
762:
747:"Applications"
738:
714:
705:
696:
687:
678:
651:
642:
632:
623:
614:
600:
575:
548:
547:
545:
542:
541:
540:
535:
530:
525:
520:
513:
510:
475:cell membranes
431:carbon dioxide
395:
392:
376:residence time
324:
321:
311:
308:
196:homogenization
163:
160:
156:cell unroofing
142:
139:
133:from inside a
114:
113:
28:
26:
19:
9:
6:
4:
3:
2:
789:
778:
775:
774:
772:
752:
748:
742:
728:
724:
718:
709:
700:
691:
682:
666:
662:
655:
646:
636:
627:
618:
610:
604:
590:on 2017-04-25
589:
585:
579:
564:
560:
553:
549:
539:
536:
534:
531:
529:
526:
524:
521:
519:
516:
515:
509:
507:
503:
499:
495:
491:
487:
482:
480:
476:
472:
468:
463:
460:
456:
452:
447:
444:
440:
436:
435:nitrous oxide
432:
428:
423:
421:
416:
412:
408:
403:
401:
391:
388:
384:
380:
377:
371:
369:
365:
364:microchannels
360:
358:
357:nanoemulsions
354:
349:
344:
340:
338:
332:
330:
320:
316:
307:
305:
301:
297:
293:
288:
283:
281:
277:
274:
270:
264:
262:
258:
254:
250:
245:
243:
237:
235:
231:
226:
224:
220:
217:
213:
209:
205:
201:
197:
193:
189:
185:
181:
177:
173:
169:
159:
157:
152:
148:
138:
136:
132:
128:
120:
110:
107:
99:
96:November 2008
88:
85:
81:
78:
74:
71:
67:
64:
60:
57: –
56:
52:
51:Find sources:
45:
41:
35:
34:
29:This article
27:
23:
18:
17:
777:Cell biology
754:. Retrieved
750:
741:
730:. Retrieved
726:
717:
708:
699:
690:
681:
669:. Retrieved
664:
654:
645:
635:
626:
617:
603:
592:. Retrieved
588:the original
578:
566:. Retrieved
562:
552:
483:
479:biochemicals
464:
448:
424:
404:
397:
389:
385:
381:
372:
361:
345:
341:
333:
326:
317:
313:
287:fluorocarbon
284:
276:vial holders
265:
256:
246:
238:
230:vortex mixer
227:
165:
144:
126:
125:
102:
93:
83:
76:
69:
62:
50:
38:Please help
33:verification
30:
538:Homogenizer
490:plant cells
457:that might
451:subcellular
368:shear rates
162:Bead method
756:2019-12-13
732:2019-12-13
671:9 November
594:2017-04-24
544:References
528:Sonication
473:, prepare
471:organelles
467:homogenize
415:ultrasonic
411:organelles
66:newspapers
486:mammalian
455:attrition
427:oxidation
362:By using
257:SoniBeast
180:suspended
151:culturing
771:Category
512:See also
494:bacteria
459:denature
443:nitrogen
420:shearing
353:liposome
304:bacteria
273:aluminum
223:aerosols
200:membrane
563:embl.de
407:enzymes
172:ceramic
147:cloning
141:Methods
80:scholar
568:19 May
506:spores
502:fungus
208:animal
204:spores
182:in an
82:
75:
68:
61:
53:
640:York.
518:Lysis
498:Yeast
413:than
348:lysed
300:algae
219:lysis
216:yeast
212:plant
176:steel
174:, or
168:glass
87:JSTOR
73:books
673:2022
570:2015
409:and
302:and
242:vial
210:and
149:and
135:cell
59:news
269:ice
206:to
42:by
773::
749:.
725:.
663:.
561:.
504:,
500:,
437:,
433:,
306:.
244:.
170:,
158:.
137:.
759:.
735:.
675:.
611:.
597:.
572:.
109:)
103:(
98:)
94:(
84:·
77:·
70:·
63:·
36:.
Text is available under the Creative Commons Attribution-ShareAlike License. Additional terms may apply.
