536:
281:. The ultramicrotome is also used with its glass knife or an industrial grade diamond knife to cut survey sections prior to thin sectioning. These survey sections are generally 0.5 to 1 μm thick and are mounted on a glass slide and stained to locate areas of interest under a light microscope prior to thin sectioning for the TEM. Thin sectioning for the TEM is often done with a gem quality diamond knife. Complementing traditional TEM techniques ultramicrotomes are increasingly found mounted inside an SEM chamber so the surface of the block face can be imaged and then removed with the microtome to uncover the next surface for imaging. This technique is called
515:/cm, can be achieved. Through the non-linear interaction of the optical penetration in the focal region a material separation in a process known as photo-disruption is introduced. By limiting the laser pulse durations to the femtoseconds range, the energy expended at the target region is precisely controlled, thereby limiting the interaction zone of the cut to under a micrometre. External to this zone the ultra-short beam application time introduces minimal to no thermal damage to the remainder of the sample.
552:
491:
528:
153:
377:
1807:
444:(SBFSEM), and are sometimes also important for light-optical microscopy. The typical thickness of these cuts is between 40 and 100 nm for transmission electron microscopy and often between 30 and 50 nm for SBFSEM. Thicker sections up to 500 nm thick are also taken for specialized TEM applications or for light-microscopy survey sections to select an area for the final thin sections.
404:
567:"knife-maker" fracturing devices. Glass knives may be used for initial sample preparations even where diamond knives may be used for final sectioning. Glass knives usually have small troughs, made with plastic tape, which are filled with water to allow the sample to float for later collection. Diamond blades may be built into such an existing trough, allowing for the same collection method.
421:
348:
1142:
713:
1851:
225:
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365:
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angles between the sample and the microtome knife, the pressure applied to the sample during the cut can be reduced. Typical applications for this design of microtome are of the preparation of large samples, such as those embedded in paraffin for biological preparations. Typical cut thickness achievable on a sledge microtome is between 1 and 60 μm.
433:. It allows the preparation of extremely thin sections, with the device functioning in the same manner as a rotational microtome, but with very tight tolerances on the mechanical construction. As a result of the careful mechanical construction, the linear thermal expansion of the mounting is used to provide very fine control of the thickness.
583:
The inclination of the knife is the angle between the knife face and the sample. For an optimal result, this angle must be chosen appropriately. The optimal angle depends upon the knife geometry, the cut speed and many other parameters. If the angle is adjusted to zero, the knife cut can often become
579:
The declination is the angle of contact between the sample vertical and knife blade. If the knife blade is at right angles (declination=90) the cut is made directly using a pressure based mode, and the forces are therefore proportionally larger. If the knife is tilted, however, the relative motion of
506:
is not required, thereby minimizing the artifacts from preparation methods. Alternately this design of microtome can also be used for very hard materials, such as bones or teeth, as well as some ceramics. Dependent upon the properties of the sample material, the thickness achievable is between 10 and
466:
The vibrating microtome operates by cutting using a vibrating blade, allowing the resultant cut to be made with less pressure than would be required for a stationary blade. The vibrating microtome is usually used for difficult biological samples. The cut thickness is usually around 30–500 μm for
575:
Prior to cutting by microtome, biological materials are usually placed in a more rigid fixative, in a process known as embedding. This is achieved by the inflow of a liquid substance around the sample, such as paraffin (wax) or epoxy, which is placed in a mold and later hardened to produce a "block"
355:
A sledge microtome is a device where the sample is placed into a fixed holder (shuttle), which then moves backwards and forwards across a knife. Modern sled microtomes have the sled placed upon a linear bearing, a design that allows the microtome to readily cut many coarse sections. By adjusting the
215:
Today, the majority of microtomes are a knife-block design with a changeable knife, a specimen holder and an advancement mechanism. In most devices the cutting of the sample begins by moving the sample over the knife, where the advancement mechanism automatically moves forward such that the next cut
164:
development, sections from plants and animals were manually prepared using razor blades. It was found that to observe the structure of the specimen under observation it was important to make clean reproducible cuts on the order of 100 μm, through which light can be transmitted. This allowed for
518:
The laser radiation is directed onto a fast scanning mirror-based optical system, which allows three-dimensional positioning of the beam crossover, whilst allowing beam traversal to the desired region of interest. The combination of high power with a high raster rate allows the scanner to cut large
562:
Planar concave microtome knives are extremely sharp, but are also very delicate and are therefore only used with very soft samples. The wedge profile knives are somewhat more stable and find use in moderately hard materials, such as in epoxy or cryogenic sample cutting. Finally, the chisel profile
388:
The flywheel in many microtomes can be operated by hand. This has the advantage that a clean cut can be made, as the relatively large mass of the flywheel prevents the sample from being stopped during the sample cut. The flywheel in newer models is often integrated inside the microtome casing. The
384:
In the figure to the left, the principle of the cut is explained. Through the motion of the sample holder, the sample is cut by the knife position 1 to position 2, at which point the fresh section remains on the knife. At the highest point of the rotary motion, the sample holder is advanced by the
411:
For the cutting of frozen samples, many rotary microtomes can be adapted to cut in a liquid-nitrogen chamber, in a so-called cryomicrotome setup. The reduced temperature allows the hardness of the sample to be increased, such as by undergoing a glass transition, which allows the preparation of
248:, which when cooled forms a solid block. The tissue is then cut in the microtome at thicknesses varying from 2 to 50 μm. From there the tissue can be mounted on a microscope slide, stained with appropriate aqueous dye(s) after removal of the paraffin, and examined using a light microscope.
566:
For ultramicrotomes, glass and diamond knives are required, the cut breadth of the blade is therefore on the order of a few millimetres and is therefore significantly smaller than for classical microtome knives. Glass knives are usually manufactured by the fracture of glass bars using special
535:
510:
The device operates using a cutting action of an infrared laser. As the laser emits a radiation in the near infrared, in this wavelength regime the laser can interact with biological materials. Through sharp focusing of the probe within the sample, a focal point of very high intensity, up to
322:
instead of a mechanical knife. This method is contact-free and does not require sample preparation techniques. The laser microtome has the ability to slice almost every tissue in its native state. Depending on the material being processed, slice thicknesses of 10 to 100 μm are feasible.
272:
Technique: after embedding tissues in epoxy resin, a microtome equipped with a glass or gem grade diamond knife is used to cut very thin sections (typically 60 to 100 nanometer). Sections are stained with an aqueous solution of an appropriate heavy metal salt and examined with a
306:) Technique: thin polymer sections are needed in order that the infra-red beam can penetrate the sample under examination. It is normal to cut samples to between 20 and 100 μm in thickness. For more detailed analysis of much smaller areas in a thin section, FTIR
475:
The saw microtome is especially for hard materials such as teeth or bones. The microtome of this type has a recessed rotating saw, which slices through the sample. The minimal cut thickness is approximately 30 μm and can be made for comparatively large samples.
448:(preferably) and glass knives are used with ultramicrotomes. To collect the sections, they are floated on top of a liquid as they are cut and are carefully picked up onto grids suitable for TEM specimen viewing. The thickness of the section can be estimated by the
258:; sections are stained and examined with a light microscope. This technique is much faster than traditional histology (5 minutes vs 16 hours) and is used in conjunction with medical procedures to achieve a quick diagnosis. Cryosections can also be used in
389:
typical cut thickness for a rotary microtome is between 1 and 60 μm. For hard materials, such as a sample embedded in a synthetic resin, this design of microtome can allow good "semi-thin" sections with a thickness of as low as 0.5 μm.
693:
372:
This instrument is a common microtome design. This device operates with a staged rotary action such that the actual cutting is part of the rotary motion. In a rotary microtome, the knife is typically fixed in a vertical position.
211:
At the end of the 1800s, the development of very thin and consistently thin samples by microtomy, together with the selective staining of important cell components or molecules allowed for the visualisation of microscope details.
587:
If the angle is too large, the sample can crumple and the knife can induce periodic thickness variations in the cut. By further increasing the angle such that it is too large one can damage the knife blade itself.
196:
The apparatus has enabled a precision in work by which I can achieve sections that by hand I cannot possibly create. Namely it has enabled the possibility of achieving unbroken sections of objects in the course of
424:
A ribbon of ultrathin sections prepared by room-temperature ultramicrotomy, floating on water in the boat of a diamond knife used to cut the sections. The knife blade is the edge at the upper end of the trough of
208:
The obscurities in the origins of the microtome are due to the fact that the first microtomes were simply cutting apparatuses, and the developmental phase of early devices is widely undocumented.
519:
areas of sample in a short time. In the laser microtome the laser-microdissection of internal areas in tissues, cellular structures, and other types of small features is also possible.
667:
543:
The selection of microtome knife blade profile depends upon the material and preparation of the samples, as well as the final sample requirements (e.g. cut thickness and quality).
175:
In 1835, Andrew
Prichard developed a table based model which allowed for the vibration to be isolated by affixing the device to the table, separating the operator from the knife.
1124:
559:
Generally, knives are characterized by the profile of the knife blade, which falls under the categories of planar concave, wedge shaped or chisel shaped designs.
441:
282:
1389:
1165:
1195:
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semi-thin samples. However the sample temperature and the knife temperature must be controlled in order to optimise the resultant sample thickness.
1897:
1907:
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the knife is increasingly parallel to sample motion, allowing for a slicing action. This behaviour is very important for large or hard samples
966:
885:
791:
168:
One of the first devices for the preparation of such cuts was invented in 1770 by George Adams, Jr. (1750–1795) and further developed by
1872:
172:. The device was hand operated, and the sample held in a cylinder and sections created from the top of the sample using a hand crank.
909:
854:
829:
627:
The
Construction of Timber, from its early growth; Explained by Microscope, and proven from Experiments, in a great Variety of Kinds
1854:
1121:
991:
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for a chosen thickness can be made. The section thickness is controlled by an adjustment mechanism, allowing for precise control.
128:
Microtomy is a method for the preparation of thin sections for materials such as bones, minerals and teeth, and an alternative to
437:
274:
136:. Microtome sections can be made thin enough to section a human hair across its breadth, with section thickness between 50
101:
depending upon the specimen being sliced and the desired thickness of the sections being cut. Steel blades are used to prepare
502:
microtome is an instrument for contact-free slicing. Prior preparation of the sample through embedding, freezing or chemical
531:
A diamond knife blade used for cutting ultrathin sections (typically 70 to 350 nm) for transmission electron microscopy
1188:
1842:
1820:
913:
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with its blunt edge, raises the stability of the knife, whilst requiring significantly more force to achieve the cut.
202:
1954:
1912:
1837:
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254:: water-rich tissues are hardened by freezing and cut in the frozen state with a freezing microtome or microtome-
1642:
1542:
1537:
748:
Loukas M, Clarke P, Tubbs RS, Kapos T, Trotz M (2008). "The His family and their contributions to cardiology".
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251:
1949:
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Serial sectioning: obtaining a continuous ribbon of sections from a paraffin block and using all for slides.
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109:. Glass knives are used to slice sections for light microscopy and to slice very thin sections for
652:
1944:
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799:
1012:"Thin Sections: A study of section thickness and physical distortion produced during microtomy"
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colors of reflected light that are seen as a result of the extremely low sample thickness.
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and tough plant matter for both light microscopy and for electron microscopy. Gem-quality
1159:
646:
904:(Histology: Cytology, general Histology, microscopial anatomy) Walter de Gruyter, 1998,
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same thickness as the section that is to be made, allowing the next section to be made.
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Occasionally, attribution for the invention of the microtome is given to the anatomist
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1332:
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1219:
292:. These microtomes have heavier blades and cannot cut as thin as a regular microtome.
122:
76:
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Botanical
Microtomy Technique: hard materials like wood, bone and leather require a
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Mikrobiologische
Vereinigung München e. V., 2006, accessed 15 February 2009
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17:
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1064:"Development of a live tissue microtome: reflections of an amateur machinist"
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205:. Several sources describe the Purkyne model as the first in practical use.
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the observation of samples using light microscopes in a transmission mode.
113:. Industrial grade diamond knives are used to slice hard materials such as
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erratic, and a new location of the knife must be used to smooth this out.
244:
Technique: tissues are fixed, dehydrated, cleared, and embedded in melted
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1707:
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1204:
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Werner
Gerabek, Bernhard D. Haage, Gundolf Keil, Wolfgang Wegner (2005):
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The cutting edge of a disposable blade for a microtome under a microscope
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Other sources further attribute the development to a Czech physiologist
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Histologie: Zytologie, allgemeine
Histologie, mikroskopische Anatomie.
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Principle of sample movement for making a cut on a rotary microtome
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as freezing tissue stops degradation of tissue faster than using a
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Handbuch der molekularen
Medizin (Handbook of molecular medicine)
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are also used for slicing thin sections for electron microscopy.
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1158:
959:(Histology : practical textbook for analytical biomedicine)
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674:. Oxford, England: The Royal Microscopical Society: 779–782.
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and does not alter or mask its chemical composition as much.
326:
Sectioning intervals can be classified mainly into either:
55:
used to produce extremely thin slices of material known as
333:
Step sections: collected at specified depths in the block.
729:. Encyclopædia Britannica Online. Encyclopædia Britannica
849:(Encyclopaedia of medical history), Walter de Gruyter,
436:
These extremely thin cuts are important for use with
1700:
1598:
1513:
1454:
1388:
1298:
1218:
928:The Theory and Practice of Histological Techniques
690:Transactions of the American Microscopical Society
156:A diagram of a microtome drawn by Cummings in 1770
1120:, WILEY-VCH Verlag GmbH, Biophotonics, S. 49–51 (
648:A Practical Treatise on the use of the Microscope
494:A conceptual diagram of laser microtome operation
467:live tissue and 10–500 μm for fixed tissue.
655:, Chapter XII (Microtomes and Microtome Knives).
194:
875:Die Entwicklung der biologischen Gedankenwelt.
442:serial block-face scanning electron microscopy
283:serial block-face scanning electron microscopy
1189:
8:
926:Bancroft, John; Stevens, Alan, eds. (1982).
900:Werner Linß, Werner Linb, Jochen Fanghänel:
686:The Development of Botanical Microtechnique.
183:
619:
617:
1196:
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877:(The evolution of the biological thought )
672:Journal of the Royal Microscopical Society
228:Microtome (C. Reichert, Vienna, 1905–1915)
1038:
651:. London: Hippolyte Bailliere. pp.
419:
368:A rotary microtome of older construction
318:, which cuts the target specimen with a
151:
105:sections of animal or plant tissues for
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27:Tool to cut fine samples for microscopy
982:
980:
978:
930:(2nd ed.). Longman Group Limited.
1062:Krumdieck, Carlos L. (January 2013).
289:
277:. This instrument is often called an
7:
1828:
429:An ultramicrotome is a main tool of
1850:
750:International Journal of Cardiology
310:can be used for sample inspection.
75:for observation under transmitted
25:
668:"An eighteenth century Microtome"
1849:
1827:
1806:
1805:
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438:transmission electron microscope
275:transmission electron microscope
232:The most common applications of
59:, with the process being termed
630:. London: The author. pp.
989:Das Schneiden mit dem Mikrotom
847:Enzyklopädie Medizingeschichte
1:
692:34, Nr. 2. 1915, S. 71–129, (
1080:10.3109/00498254.2012.724727
762:10.1016/j.ijcard.2006.12.070
555:Profiles of microtome knives
314:A recent development is the
185:Beschreibung eines Mikrotoms
1116:Holger Lubatschowski 2007:
961:. Springer, Wien/New York.
694:PDF-Version of the article)
1971:
483:
459:
396:
190:Description of a Microtome
29:
1801:
1338:Hemispherical photography
1287:
67:, microtomes are used in
51:, meaning "to cut") is a
1898:Machine and metalworking
1643:Lombard Steam Log Hauler
994:10 November 2009 at the
399:Frozen section procedure
252:Frozen section procedure
30:For the moth genus, see
1908:Measuring and alignment
1166:Encyclopædia Britannica
1019:J Biophys Biochem Cytol
1010:Peachey Lee D. (1958).
203:Jan Evangelista Purkyně
47:, meaning "small", and
1169:(11th ed.). 1911.
1137:10.1002/opph.201190252
684:Gilbert Morgan Smith:
645:Quekett, John (1848).
576:which is readily cut.
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450:thin-film interference
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73:preparation of samples
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304:infrared spectroscopy
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160:In the beginnings of
155:
1873:Cutting and abrasive
1855:WikiProject Forestry
1127:19 July 2011 at the
954:Gudrun Lang (2006).
547:Design and cut types
260:immunohistochemistry
1398:Aerial firefighting
1031:10.1083/jcb.4.3.233
871:Ernst Mayr (2002).
624:Hill, John (1770).
270:Electron Microscopy
111:electron microscopy
1738:Hydraulic debarker
1713:Firewood processor
666:Anonymous (1910).
557:
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427:
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230:
192:), Wilhelm wrote:
170:Alexander Cummings
158:
1932:
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1743:Log house moulder
1269:Tree planting bar
1244:Groasis Waterboxx
1160:"Microtomy"
968:978-3-211-33141-5
887:978-3-540-43213-5
320:femtosecond laser
16:(Redirected from
1962:
1955:Cutting machines
1853:
1852:
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1809:
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1788:Wood-drying kiln
1723:Forestry mulcher
1673:Washington Winch
1575:Portable sawmill
1390:Fire suppression
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802:on 25 April 2009
798:. Archived from
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351:A sled microtome
290:sledge microtome
187:
180:Wilhelm His, Sr.
162:light microscope
130:electropolishing
107:light microscopy
43:(from the Greek
32:Microtome (moth)
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1118:Laser Microtomy
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182:(1865). In his
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86:Microtomes use
63:. Important in
61:microsectioning
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1680:Steam donkey
1565:Lumber edger
1550:Crosscut saw
1495:Log splitter
1420:Fire flapper
1375:Tree caliper
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1343:Inclinometer
1328:Cruising rod
1277:(Tuley tube)
1275:Tree shelter
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800:the original
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756:(2): 75–78.
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1585:Two-man saw
1500:Marking axe
1481:(shake axe)
1380:Wedge prism
1353:Rangefinder
1313:Angle gauge
1300:Mensuration
1233:Caulk boots
1068:Xenobiotica
796:msn Encarta
792:"Histology"
134:ion milling
83:radiation.
1939:Categories
1811:Categories
1633:Helicopter
1474:Brush hook
1308:3D scanner
1281:Tree spade
1254:Pottiputki
1074:(1): 2–7.
609:References
603:Microscope
571:Sectioning
484:See also:
440:(TEM) and
397:See also:
308:microscopy
234:microtomes
69:microscopy
1945:Histology
1843:equipment
1821:equipment
1773:Tree tyer
1753:Log flume
1733:Hand hook
1663:Skid cone
1658:Pike pole
1653:Pickaroon
1638:Log truck
1628:Harvester
1618:Forwarder
1608:Cant hook
1442:(rakehoe)
1425:Fire rake
1415:Driptorch
1370:Relascope
1365:Microtome
1264:Seed trap
1212:equipment
1088:0049-8254
770:0167-5273
598:Histology
462:Vibratome
456:Vibrating
285:(SBFSEM).
242:Histology
197:research.
41:microtome
1878:Forestry
1868:Cleaning
1748:Log pond
1708:Denailer
1623:Go-devil
1560:Head saw
1533:Chainsaw
1469:Broadaxe
1464:Billhook
1435:Helitack
1240:(hoedag)
1205:Forestry
1125:Archived
1096:23009272
1049:13549493
992:Archived
806:18 March
778:17433467
733:24 March
592:See also
504:fixation
264:fixative
256:cryostat
246:paraffin
81:electron
71:for the
57:sections
1903:Masonry
1893:Kitchen
1833:Commons
1758:Machete
1668:Skidder
1600:Logging
1590:Whipsaw
1570:Polesaw
1555:Dragsaw
1528:Bucksaw
1523:Bow saw
1485:Hatchet
1446:Pulaski
1249:Mattock
1104:6108637
1040:2224471
701:3221940
148:History
96:diamond
65:science
49:temnein
1913:Mining
1883:Garden
1778:Tsakat
1685:Yarder
1648:Peavey
1490:Labrys
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425:water.
360:Rotary
343:Sledge
99:blades
45:mikros
1918:Power
1838:tools
1816:tools
1701:Other
1690:swing
1580:Resaw
1403:DC-10
1358:laser
1323:Chain
1208:tools
1100:S2CID
1015:(PDF)
697:JSTOR
500:laser
480:Laser
338:Types
236:are:
119:teeth
92:glass
88:steel
77:light
1888:Hand
1515:Saws
1479:Froe
1456:Axes
1408:UAVs
1210:and
1092:PMID
1084:ISSN
1045:PMID
963:ISBN
906:ISBN
882:ISBN
851:ISBN
826:ISBN
808:2009
774:PMID
766:ISSN
735:2009
688:In:
498:The
300:FTIR
132:and
115:bone
1133:doi
1131:).
1122:PDF
1076:doi
1035:PMC
1027:doi
857:, (
832:, (
758:doi
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704:doi
653:306
471:Saw
302:or
94:or
79:or
1941::
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513:TW
144:.
142:μm
138:nm
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39:A
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34:.
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Text is available under the Creative Commons Attribution-ShareAlike License. Additional terms may apply.
