This short article tells how to check any image for empty magnification. It also helps to assess images with focusing issues, because wrong focus has similar result: lack of small (1-2 pixels) details.
Method
Method is based on so called "Wavelet decomposition". This mathematical concept stands for splitting an image into scale planes, where each plane contains details of specific size: 1 pixel, 2 pixels, 4 pixels and so on (these are powers of 2, starting from 0).
Having all details sorted by size and split into planes, it's possible to check, if planes, supposed to contain smallest details, are empty or not. Technically, it's important to remember, that any image also contains certain amount of noise, originating from camera sensor, for example. Therefore, smallest details plane will always contain something, but it could be just noise. This plane can also contain much less details than others. It means, that optical system works somewhere on its limit, but not beyond it.
Software
This method utilizes freeware cross-platform image editor, called GIMP (don't download it from there if you are not an experienced programmer). There are both ready to use binary distributions for different operation systems (Windows, Mac, Linux) and source code available.
You will also have to have Wavelet decompose plugin installed.
Windows and Mac OS users can obtain installations from Partha's place website. It has Wavelet decompose plugin already incorporated into distribution file, so you don't need to compile it.
Install GIMP using one of distributions, make sure it has Wavelet decomposition plugin and you are good to go.
Instruction
It doesn't have any sense to assess combined stacks, because stacking software obviously distorts the result.
Here, I will use cropped portion of spider skin photo taken with lens, mounted on bellows:

Open the image in GIMP, go to Filters - Generic - Wavelet decompose ... menu:

Technically, you need only a couple of fine details planes, but it makes sense to generate five planes, containing details from 1px to 16px size. To do that, use these settings:

You will get six new layers (five planes plus "Residuals"). Turn off all of them, except "Wavelet scale 1" - finest details layer. It will look like this:

Probably, you can barely see anything there (in this exact case, optical system works on its limit, therefore, there are not so much details). Let's check, if larger details layer contains more.
To do that, turn off "Wavelet scale 1" and turn on "Wavelet scale 2":

Second layer obviously reveals more details. But let's get back to the finest layer (turn off "Wavelet scale 2" and turn on "Wavelet scale 1"). Using conventional image enhancement methods, it's possible to make details from "Wavelet scale 1" more pronounced and easier to assess.
To do that, select "Wavelet scale 1" layer in list (click on it), go to Colours - Levels... menu and use Auto button to apply automatic channel equalization to currently selected layer:

Here is the resulting enhanced image:

As you can see, it does contain a lot of noise, but there are also details of legs. It means, this image doesn't suffer from "empty magnification" effect.
Notes
Adobe Photoshop or ImageJ (Fiji), widely used for image processing, do not have any similar tools. There are certain ways to use this method there, utilizing Fast Fourier Transform or KPT Equalizer plugin for Photoshop (no longer supported and incompatible with 64-bit version of Photoshop), therefore I will unlikely be able to answer questions regarding usage of this method with software, other than GIMP.
Wavelet decomposition can be effectively used for detail enhancement. Increasing the contrast of any detail plane makes details of corresponding size more pronounced in image after fusing the layers back together. Local contrast manipulation (such as applying Gaussian blur to selected empty areas of "Wavelet scale 1" layer) is an effective method of noise reduction.