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What are close-up and macro ? ‹ Close-up photography - photographic images taken close to the subject typically with magnifications of ~0.1X to 1X. Close-up and ‹ Macro (or micro) photography - images taken with a magnification of 1X (or full size) , i.e., the size of the object is reproduced at the same size on the camera’s Macro Photography sensor or film . ‹ Extreme macro photography - images taken with Michael K. Miller magnifications higher than 1X to ~15X CCOR July 12, 2016 ‹ Photomicroscopy or photomicrography - images taken with a variety of microscopes at magnifications between ~10X and >1,000,000X. ‹ Macro a prefix from the Greek for long or large

Field Ion Microscopy ~5 million X 1 each spot is an atom 2

1.5 mm, 0.06 in Small objects Overview of the Different Techniques

90% The tools that you require to of near minimum focus distance frame photograph small objects depend on the physical size of the object and its Point and shoot cameras in macro mode distance to the camera. Macro lenses

Magnification is size of the image Close-up lenses compared to the size of the subject Reversed lenses

~7.4X magnification Extension tubes and bellows 24 x 36 mm sensor Macro photography was first practiced somewhere between 1835 and 1839 by Twin (or stacked) configuration Englishman William Henry Fox Talbot (1800–1877). However, the term photo-macrograph (macro) was later proposed in 1899 by W. H. Walmsley for close-up images with less than 10 diameters magnification. 1X 5X 10X 15X Practicalities The German Fritz Goro (Goreau 1901-86) is sometimes credited for inventing Lighting at small working distances macrophotography, making visible the world that lies between the microscope Positioning subject and the naked eye . Resolution verses DOF Early macro SLR lenses: 1955 Kilfitt Makro-Kilar 4 cm f/3.5 for Exakta 35mm SLRs Focus Stacking 4 1959 13.5cm f/4 Q: 1961 Nikon 5.5cm f/3.5 Micro 3 4

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Visualizing magnification Regular camera + telephoto lens

8X 3 x 4.5 mm near minimum focus distance 8X The minimum (or close) focus distance is the shortest distance from 4X 4X 6 x 9 mm the sensor plane to the subject at which a lens can focus. 2X 2X 12 x 18 mm The working distance is the distance from the front of the lens to the subject. Hand held Wide postage stamp Telephoto lenses, either handheld or on 1X 24 x 36 mm FF Sensor a tripod, can enable capture of images 1X when the subject is sensitive to close US quarter is approach or is moving. 0.955 inches Once you know the size of your or 24.26 mm subject then you can choose Nikon Lenses W.D. inch Mag. in diameter Height 70-200 f/4 30.5 0.27X Dime is of the optimum equipment to 0.705 inches 70-200 f/2.8 45.3 0.25X or 17.91 mm subject record its image. in diameter 300 PF f/4E 47.6 0.24X A nickel is 2:1 = 2X – twice size on sensor © Michael K. Miller 1/16 inch or 80-400 f/4.5-6.3 59.0 0.20X

1.58 mm OnFull Frame body Nikon D7000 + 70-200mm lens @ 180mm, f/5.6 1:1 = 1X – same size on sensor 200-500 f/5.6E 79.7 0.22X thick 0.5X 1:2 = 0.5X – half size on sensor Olympus 300 f/4 45.3 0.48X, 0.67X Extension tubes can be used to reduce the (MFT) with 1.4X TC minimum focus distance (useful for long lenses) 1 inch = 25.4 mm 5 6

© Michael K. Miller © Michael K. Miller with 80-400 mm lens @ 400 mm, f/8, 1/1000s, 800 ISO Nikon D810 with 300 mm lens @ f/8, 1/200s, 400 ISO, handheld handheld in boat 7 8

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Point and shoot camera in “Macro” mode Pre-Columbian Gold Museum in San José, Costa Rica 40 mm Most P&S Canon G7X in Macro mode cameras have a “so-called” Most P&S macro mode cameras have to enable a “so-called” photographing macro mode small objects. to enable photographing small objects.

Cropped & hand held ~40 mm high on 1” sensor sensor at minimum size is 8.8 x 13.2 mm focus distance in Canon G7X © Michael K. Miller Therefore, maximum magnification of the G7X is 8.8/40 = 0.22X Canon PowerShot S100 in Macro mode, handheld Working distance at this magnification is only ~1.5 inches. focal length 24 mm (35mm equiv), f/2, 1/30 s, 160 ISO, no flash

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$500 Red-eyed Tree Frog, Costa Rica Macro lens Macro lenses have their optical formula optimized for close-up photography with magnifications up to 1X (1:1). Macro lenses are designed with a flat focus field (rather than the normal curved field) to provide a uniformly sharp image across the entire frame. The magnification increases as the focus distance decreases to the minimum focus distance. Magnification Focus distance 13 mm

Olympus M.ZUIKO DIGITAL © Michael K. Miller 60mm f/2.8 ED Macro Effective focal length = 120 mm Canon Powershot G7X in Macro mode - handheld increased due to 2X crop factor 1X focal length 35 mm (35 mm equiv.), f/3.2, 1/40 s, 125 ISO, no flash Sensor size 13 x 17.3 mm 11 12

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$250 - 40mm $1800 - 200 mm Macro lenses Macro lenses LIVE subjects All true macro lenses can achieve a ° The maximum magnification, 1X, is at the minimum 1X magnification on the sensor focus distance. irrespective of the focal length of Use by setting focus at 1:1 distance and move camera along the lens . the optic axis to achieve focus Macro lenses are available in focal ° Some macro lenses suffer from shortening of the lengths from ~15 mm to ~200 mm. focal length near the close focusing distance, The longer focal lengths have a i.e., the magnification changes dramatically in the longer working distance but are range (0.5X to 1X) 1:2 to 1:1, For example, Nikon AF micro-Nikkor 200 mm is ~102 mm at 1X heavier and more expensive Canon EF 100 mm is ~93 mm at infinity and ~64 mm at 1X more lighting options + less shadows This change in magnification can make framing difficult . less intrusive on live creatures They can also be used for regular 1X ° Effective f-number increases with magnification, m: photography and will focus to Neff = N(1+ m) infinity. ° Use tethered Live View or a right-angle finder Olympus OM-D E1, 60 mm macro Macro lenses typically include the if necessary @ f/7.1, 1/500s, ° Useful for copying slides and as a portrait lens ISO 200, reproduction ratio 1:2, 1:1 etc. handheld above the distance scale. 13 14

Nikon 200 mm f/4 macro lens Canon 65 mm 1-5X Macro Lens http://www.pierretoscani.com/echo_focal_length.html can achieve magnifications similar to extension tubes/bellows $1500 MP-E f/2.8

Manual focus lens extends with higher magnifications (not an IF lens)

Closest Focusing Working distances Distance mm inch 240 mm, 9.6 inches 1X 101 4.0 (from film plane to 2X 63 2.5 3X 51 2.0 subject ) 4X 44 1.8 The Nikon 200 mm f/4 macro lens changes focal length dramatically near 5X 41 1.6 the minimum focus distance. 15 16

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Other methods Lensbaby and cellphones ~$400 ~$70

Macro lens sets to clip The Composer Pro Macro Pack includes the onto various cellphones. Composer Pro with Sweet 50 Optic, Optic, Very close focus distance. Macro Converters BLIMPS microlenses: The Macro Converters sit in between the optic and kickstarter.com/projects/blips/ © Michael K. Miller lens body and allow for focus between 5" and 8.13" blips/description away from your subject. Nikon D800 + 200 mm f/4 Macro mode; 100 ISO, 3 sec 1X 17 f/20 used due to the depth-of-fieldrequired Sensor size 24 x 36 mm (FF) 18

Close-up (or supplementary) lens Equivalent to adding reading glasses ~$15 - 75 for set of 3 or 4 Can be stacked (add together the diopters) Attaches to filter thread on lens wikimedia.org

Close-up filters come in different powers from +1 to +10 diopters diopter power = 1000 /focal length of the lens (in mm) Therefore: +1 = 1000 mm, +2 = 500 mm, +4 = 250 mm, +10 = 100 mm = Magnification increases with longer focal lens lenses and/or higher diopter close-up lenses . © Michael K. Miller For a 50 mm lens and +4 diopter lens gives a magnification of 0.2X D800 + 200 mm f/4 Macro @ f/8, 1/250 s, 81 frames, 400 ISO 1X For a 300 mm lens and +10 diopter lens gives a magnification of 3X 19 The stated focal length of lenses are approximate and can have an error up to ~±5%. 20

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Achromatic Close-up lenses $75-$90 $75-$150 250D 500D +4 diopters +2 diopters Canon 250 D and 500 D Focal length 250 mm Focal length 500 mm close-up lenses have Suitable for 30-135mm Suitable for 70-300mm 2 achromatic elements lenses lenses to correct chromatic aberrations.

For a 300 mm lens with the 500D at ∞ focus at closest focus An achromatic Working 20.25” 515 mm 13 3/8” 340 mm lens is able to distance emit, transmit, or receive light without 0.57X 0.86X separating it Effective focal length into colors of 300 mm © Michael K. Miller lens = 230 mm 0.8X D800+200 mm f/4+Canon 500D @ f/8, 72 frame stack, 1/250s, 100 ISO Higher magnifications are possible if the is focused closer than its ∞ setting 21 22

Achromatic Close-up lenses Raynox DCR-250 and DCR-150 have 2 groups and 3 elements Adapter to fit on 52-67 mm lens filter ring 35 mm 49 mm thread

Coated high-index optical glass raynox.co.jp 43 mm plastic thread N f- 3X @ MFD Lens Diopter WD mm number w 200 mm lens 200 mm* 8 mm $49 DCR-150 +4.8 208 ≈6 145-208 mm macro lens $58 DCR-250 +8 125 ≈3.5 99-125 mm + DCR-250 Can be stacked for higher magnifications © Michael K. Miller 24x36mm sensor *Lenses change focal length near macro magnifications/distance 3X = 8x12 mm 1.6X For example, Nikon AF micro-Nikkor 200 mm is ~102 mm at 1X Nikon D800+105 mm f/2.8 @ 1:1+Raynox DCR250 @ f/32, 13s, 100 ISO Canon EF 100 mm is 93 mm at infinity and 64 mm at 1X 23 24

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Microscope lens/ Extension tubes infinity objectives 1 or more fit between the camera body and the lens Nikon CFI Plan 10x/0.25NA - 10.5mm WD Objective ~$120 for a set of 3 auto tubes Specific to a camera mount for extreme macro focus stacking Requires a 52 mm Nikon CFI Plan adapter plus a step-up ring to fit No optical components onto the filter thread size of the Can be used individually or in sets tube, i.e., a non-macro telephoto Extension 50 mm Stops CFD lens . auto tubes with electrical mm lens lost mm Best with prime lenses, but contacts recommended 5 to 10 X Tube lens none 0.15X 0 450 zooms at long end can also work 12 0.24X 0.31 137 wikimedia.org Magnificat = Working Distance only 10.5 mm 20 0.40X 0.49 107 Tube lens must be focused at infinity with wide open Shorter focal length lenses and/or longer 36 0.72X 0.78 75 Distance between the lens and the objective matters extensions give higher magnification 12+20 0.64X 0.71 81 100 mm tube lens = 5X, 200 mm lens = 10 X, 300 mm lens = 15 X There is a significant loss of light. 12+36 0.96X 0.97 61 Also reduces working distance and range of focus 20+36 1.12X 1.08 54 Infinity objectives form a virtual image at infinity; New CFD ≅ f(cfd*f + cfd*ext – ext*f)/(cfd*ext – ext*f + f*f) 12+20+36 1.36X 1.24 46 the tube lens takes these parallel rays and focuses them as a real image on the sensor. 25 1:1 Macro 50 mm lens + 50 mm extension 100 mm lens are half these values 26

Bellows G7 fits between the body and the lens sensor 7.2 X 5.3 mm

$825 for Canon $1001 for Nikon WD Novoflex Auto Bellows for Canon EOS 37 Bellows are flexible and continuously adjustable extension tubes (from 40 ( 0.8X ) - 192 mm ( 3.8X )) that mm allow precise extensions and magnifications to be set. novoflex.com Bellows can also include a tilt-shift mechanism. Mag Often used together with extension tubes and reversed lenses for even higher magnifications . 1.4X FotodioX Macro Bellows for Nikon F or Canon EOS: $50 Extensions from 40 - 150 mm ( 0.8X to 3X with 50 mm lens ) 27 Nikon D800 + 50 mm f/1.4 + 68 mm extension @ f/16, 2s, 100 ISO 28

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Reversed lens Aperture control for Reverse ring adapter fits between body and reversed lens auto focus reversed lens Lens bayonet Focus at ∞ Not to scale An inexpensive ~$12 Aperture Control Filter attaches to and protects the rear of the lens and can reduce glare in lens like a reverse ring adapter lens hood allows a lens to be mounted on a camera in the reversed position. Bayonet specific ~$25 to camera Manual lenses of any mount make preferably with Lens filter thread Aperture ring an aperture ring can Reverse ring adapter be used. Camera is operated in manual (M) mode

Adds a filter tread Auto focus lenses require the aperture to controlled manually Male thread as it is no longer controlled by the camera Bayonet specific to camera mount Filter thread specific to lens 29 30

Reversed lens -Magnification Reversed lens -Magnification Magnification cannot be calculated for compound (thick) lenses 16-35 mm at infinity focus 16 mm 4.6X 20 mm 4X 24 mm 3.3X 22 mm 28mm 2.9X 35 mm 2X ~1.1X 13 mm ~1.8X primes WD @ CFD 50 mm 1X, 23 to 30mm 85 mm 0.36X 50 mm f/1.2 lens at f/4 28 mm f/2.8 lens at f/4 W.D. 86 mm, 3.4 inch W.D. 53 mm, 2.1 inch Wide angle lenses give the highest magnification Wide angle lenses give the highest magnification Exact magnification depends on optical formula of each lens Inexpensive but often get soft edges at higher magnifications and lens focus distance 31 32

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14 mm Stacking Lenses/Macro Coupler 0.55 inch primary dia. lens reversed lens

~$10

wikimedia.org

Couplers often have the same size male threads on both sides. Use step-up or coupler step-down rings to match to lenses. Use Aperture Control Filter for AF lenses This configuration consists of a primary longer focal length lens, a macro coupler, and a reversed shorter focal length lens. The coupler screws into the filter rings on both lenses. = Examples: 50mm + reversed 50 mm = 1X; 2X 300 mm + reversed 20 mm = ~15X Nikon D800 + reversed 28 mm f/2.8 @ f/16, 1/20s, 100 ISO 33 34

Stacking Lenses/Macro Coupler

Vignetting at the corners of the frame can occur with stacked lenses due to the aperture of 1.6 mm the reversed lens. 0.063 in To minimize this effect, set the focus of the primary lens to its 15X closest focusing distance. Sensor size 24 x 36 mm Nikon D800 with AF-S 300 mm f/4 + reversed AF-S 20 mm Working distance ~1.3 inch (33 mm) Lighting: tactical flashlight focused on ruler © Michael K. Miller Nikon D800 300 mm + reversed 50 mm f/1.4 @ f/8, 1/20s, 100 ISO 6X 35 36

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Methods to get 1:1 (1X) magnification Reduction of working range 1. Macro lens at close focus distance subject 2. Lens + extension tubes of the same extent as the Min. focus focal length of the lens, e.g., 50 mm lens with 50 distance infinity Close-up lens mm extension subject When additional optical Max. focus 3. Stacked lenses of the same focal length, e.g., a 50 distance elements are inserted mm primary with a 50 mm reversed lens. Min. focus before or after the distance primary lens, the 4. Reversed 50 mm lens Min. focus camera has a much distance subject 5. Lens with close-up lens of the same focal lengths, Max. focus reduced range of focus e.g., a 200 mm lens and a +5 diopter (200 mm) distance and can no longer focus close-up lens Extension working at infinity. tubes distance

In some configurations, the working distance can be less than 10 mm

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Lighting the subject Light sources: flash As the working distance in macro can be very small, the ‹ On-camera flash are blocked by the lens. lens blocks much of the ambient light and also the on- ‹ Speedlights have a controllable range that is camera flash that would normally reach the subject. much longer (~0.6-20 m/2-66 ft) than the Longer focal length macro lenses give more working distance. typical macro working distance + too much working distance power. Nikon R1 macro flash 200 mm ‹ Reduce power by using manual flash Up to 4 speedlights that can be settings. working Back positioned around the ring distance lighting - ‹ A ring light goes directly around the lens and Nikon This limits the creativity for 60 lightpads is aligned with the axis of your lens resulting SB 200 artistic lighting the subject. mm with in nearly shadowless lighting but may show +windshield diffusers rings in the eyes of insects and be dimmer in 1-1500 the centre of the image. lux 6000K ‹ Two (or more) separate flash heads on either side of the lens giving a more 3D lighting effect. The SW-11 modifiers can ∑ Can be used to freeze the subject. redirect and diffuse the light

540 lux 39 ~250 flashes per charge 40

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Tactical flashlight Light sources: LED

DO NOT 130 - ~4000 lux Very bright TRY THIS AT HOME Lume Cube 1 to 1500 lumen SEVERE EYE AND/OR Light guides SENSOR with colour DAMAGE filters POSSIBLE Relatively inexpensive LED Panels and high intensity LED (tactical) flashlights with continuous light output up to 4000 lux are available. Some have adjustable color temperatures ~(3000-6000K) and intensity. Some LEDs have a blue tint – adjust White Balance accordingly. © Michael K. Miller 8X D800 + 200 mm + reversed 16-35 mm @ 35 mm, f/8, 1/20s, 100 ISO WARNING: DO NOT VIEW LED OR SHINE IN PEOPLE OR ANIMALS EYES 41 42

Fleabane “The small Positioning subject in relation to camera yellow disk At macro magnifications , $650 for pair flowers bloom from the vibration and accurate outside in. positioning are major The white concerns. reallyrightstuff.com "petals" are ray flowers.” Therefore, sturdy tripods, Kris Light ball heads , and good camera technique is essential. The MUP (mirror lock up) Note scales for focus stacking setting and a remote cable release are often used. Focus is normally achieved by moving A stable mount for the the camera with respect to the subject. 0.6X subject is also required. Lab jacks and/or macro 2- or 3-axes focusing © Michael K. Miller Moving subjects can be rails are often used for positioning the Nikon D800 + 200 mm f/4 + Canon 500D @ f/8 stack, 1/250s, 100 ISO specimen and focusing. frustrating. Single Lume Cube LED light – Auto WB 43 44

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Resolution v. v. Light Increasing DOF with Focus Stacking Higher bars better Focus stacking (or blending/Depth of Field (DOF) stacking) enables the DOF of Olympus M.ZUIKO 60mm f/2.8 ED Macro an image to be increased by combining multiple images taken at slightly DLA = 6.78*1.61 = 10.9 different focus settings or subject-to-camera distances . Allows non-planar and subjects not parallel to sensor to be photographed.

Corrected for sensor crop factor DLA Diffraction limited aperture – sensor pitch in microns X 1.61 smaller The choice of the aperture setting is a compromise between resolution/diffraction effects , depth-of-field (DOF) and amount of light required. Macro lenses are similar to other lenses in that the resolution typically increases until the refraction limited is reached at small . The DOF increases at smaller apertures and is distance dependent . Light increases with larger apertures and longer exposures Single frame at f/8 focus stack of 72 frames at f/8 The DOF is extremely small at macro working distances Most often used for macro photography but can be use for other subjects, such as landscapes. Simplest to place specimen parallel to sensor plane (parallelize the subject) DOF defines the optimum step size for focus stacking - >20% overlap. In depth how to tutorial is available on the CCOR website 46 45

Image capture for focus stacking A series of images at different focus or distances are captured. How many frames and optimum step size Example below is a ruler set at 45° to focal plane of camera Some stacking software will suggest and set the step size for the 1 2 3 camera lens or the focusing rail between the frames 4 Use at least a 20% overlap. Step size = 2 c n (m+1)/( m x m) Steps can be extremely thin when c = circle of confusion very close to subject n = f-number Thinner than a credit card (0.76 mm) 5 @ 1X = 0.45 mm with a 20% overlap 6 m = magnification 7 @10X = 0.025 mm with a 20% overlap 8 1 After capture, a focus stacking program is used to Out-of-focus bands 2 automatically align and choose the in-focus regions 3 of each image to combine into the final image. (known as focus banding ) 4 of can be observed in the 5 The shape of the in-focus regions depends on the depth profile of the subject. focus stacked image when 6 the step size is too small 7 The output may be a single jpg, tiff, or dng file of 8 the stacked image. 47 48

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Focus Stacking methods DOF 4” or 1. Turn the lens focus ring manually by hand 100 mm 2. Automatically step the focus motor inside your lens with software, e.g., ControlMyNikon™, CamRanger™, Helicon Remote ™, digiCamcontrol, etc. ‹ Must have a compatible lens with a computer chip 3. Move the camera (or subject) with a focus rail, or automatically with StackShot™ Macro rail package (cognisys-inc.com) ‹ Simplest to tether the camera to a computer. $550-850 ‹ Very time-consuming. ‹ Subject or camera must not move ‹ Optimum with extended distances and when shooting at 1:1 magnifications, i.e., when at end of focus range (no + focus may be available) Stacking software is then required to combine the images. $115 $89 See tutorial $200 $189 on CCOR $240 $289 website See en.wikipedia.org/wiki/Focus_stacking for other software Short stack – up 20 frames: Tradition stack – up to hundreds of frames Clean sensor before starting . © Michael K. Miller sensor spots will appear as dark lines stuck pixels will appear as coloured lines. D800 + 200 mm Macro @ f/8, 148 frames @ 1/250s, 400 ISO 49 50

Setup for Stackshot™ operation Parameters for the Depth Map method “Pyramid ” (Pmax) method has NO adjustable parameters Camera’s USB port “Depth map ” (Dmap) method finds the source image where the sharpest pixel is located and creates a "depth map" from this information. Radius and Smoothing parameters USB port 1 The Radius Parameter : Radius is the most important parameter. Start from the default USB port 2 value (8 HF/10 ZS) and then set it to its minimum (1) and observe the changes. Then Shutter release increase the value to get rid of any noise or artifacts, particularly halos along the edges. X If your image has fine details and thin lines, a low radius level (3-5) will probably yield the 110V Power best results, although you will probably get more noise and a halo effect. If there is a halo, try increasing the radius to minimize halo, then stop to preserve as much detail as possible. AC adapter The Smoothing Parameter : Low smoothing produces a sharper image, but the transition areas may have some artifacts. High smoothing will result in a slightly blurry image without any visible transition areas. USB power Start with the default smoothing (4 HF/5 ZS). For more detail, decrease the smoothing AC power adapter for camera eliminates the need to change batteries value; if get too much noise and too many artifacts, increase the value.

Minimum step size is 0.002 mm over the 100 or 200 mm travel of the rail. HF = Helicon Focus; ZS Zerene Stacker (Options> Preferences> Dmap settings Connect USB cables directly from computer to camera – no USB hubs. 51 52

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Popular focus stacking algorithms Some lens and reconstruction artefacts

Halos around edges of objects and in Coloured straight lines running the out-of-focus background regions perpendicularly to the edge(s) of the frame

“Pyramid ” (Pmax) method is very good at “Depth map ” (Dmap) method does a finding and preserving detail even in low better job keeping the original contrast or slightly blurred areas. smoothness and colors, but it's not as It gives good results in complex cases good at finding and preserving detail. (intersecting objects, edges, deep stacks) but Images must be in order . increases noise, contrast and glare. It can alter Substack slabbing for troublesome colors. regions or to overcome large file size/computer memory issues. Strings of dots – sometimes replicated Dmap reconstructions can introduced noise 53 similar to sensor spots and stuck pixels 54

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